Quinolinone compounds as 5-HT4 receptor agonists

ABSTRACT

The invention provides novel quinolinone-carboxamide 5-HT 4  receptor agonist compounds. The invention also provides pharmaceutical compositions comprising such compounds, methods of using such compounds to treat diseases associated with 5-HT 4  receptor activity, and processes and intermediates useful for preparing such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.11/365,320, filed Mar. 1, 2006; now U.S. Pat. No. 7,446,114 B2, whichclaims the benefit of U.S. Provisional Application No. 60/658,007, filedon Mar. 2, 2005; the entire disclosures of which are incorporated hereinby reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention is directed to quinolinone-carboxamide compounds which areuseful as 5-HT₄ receptor agonists. The invention is also directed topharmaceutical compositions comprising such compounds, methods of usingsuch compounds for treating medical conditions mediated by 5-HT₄receptor activity, and processes and intermediates useful for preparingsuch compounds.

2. State of the Art

Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter that iswidely distributed throughout the body, both in the central nervoussystem and in peripheral systems. At least seven subtypes of serotoninreceptors have been identified and the interaction of serotonin withthese different receptors is linked to a wide variety of physiologicalfunctions. There has been, therefore, substantial interest in developingtherapeutic agents that target specific 5-HT receptor subtypes.

In particular, characterization of 5-HT₄ receptors and identification ofpharmaceutical agents that interact with them has been the focus ofsignificant recent activity. (See, for example, the review by Langloisand Fischmeister, J. Med. Chem. 2003, 46, 319-344.) For example, 5-HT₄receptor agonists are useful for the treatment of disorders of reducedmotility of the gastrointestinal tract. Such disorders include irritablebowel syndrome (IBS), chronic constipation, functional dyspepsia,delayed gastric emptying, gastroesophageal reflux disease (GERD),gastroparesis, post-operative ileus, intestinal pseudo-obstruction, anddrug-induced delayed transit. In addition, it has been suggested thatsome 5-HT₄ receptor agonist compounds may be used in the treatment ofcentral nervous system disorders including cognitive disorders,behavioral disorders, mood disorders, and disorders of control ofautonomic function.

Despite the potential broad utility of pharmaceutical agents modulating5-HT₄ receptor activity, few 5-HT₄ receptor agonist compounds are inclinical use at present. One agent, cisapride, that was utilizedextensively for treatment of motility disorders of the gastrointestinaltract was withdrawn from the market, reportedly due to cardiac sideeffects. Late stage clinical trials of another agent, prucalopride, havebeen suspended.

Accordingly, there is a need for new 5-HT₄ receptor agonists thatachieve their desired effects with minimal side effects. Preferredagents may possess, among other properties, improved selectivity,potency, pharmacokinetic properties, and/or duration of action.

SUMMARY OF THE INVENTION

The invention provides novel compounds that possess 5-HT₄ receptoragonist activity. Among other properties, compounds of the inventionhave been found to be potent and selective 5-HT₄ receptor agonists.

Accordingly, the invention provides a compound of formula (I):

wherein

R¹ is hydrogen, halo, or C₁₋₄alkyl;

R² is C₃₋₄alkyl or C₃₋₆cycloalkyl;

a is 0 or 1;

Z is a moiety of formula (a):

wherein:

b is 1, 2 or 3;

d is 0 or 1;

X is carbon and Q is selected from -A-, -A(CH₂)₂N(R⁴)—, and—S(O)₂(CH₂)₂N(R⁴)—;

or X is nitrogen and Q is selected from —S(O)₂CH₂C(O)—, —SCH₂C(O)—,—OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, -A(CH₂)₂—,

G is W and c is 0, wherein W is selected from —N{C(O)R⁹}—,—N{S(O)₂R¹⁰}—, —N{C(O)OR¹²}—, —N{C(O)NR¹³R¹⁴}—, —N{S(O)₂NR¹³R¹⁴}—,—N{R¹⁶}—, —S(O)₂—, —O—, and —S—; provided that when G is W, c is 0, andb is 1, then X is carbon;

or G is carbon, c is 1, and Y is a moiety of formula (b):

wherein:

e is 0 or 1;

W′ is selected from —N(R⁸)C(O)R⁹, —N(R⁸)S(O)₂R¹⁰, —S(R¹¹)(O)₂,—N(R⁸)C(O)OR¹², —N(R⁸)C(O)NR¹³R¹⁴, —N(R⁸)S(O)₂NR¹³R¹⁴, —C(O)NR¹³R¹⁴,—OC(O)NR¹³R¹⁴, —C(O)OR¹², —OR¹⁵, and —N(R⁸)R¹⁶; provided that when X isnitrogen, e is 0, and W′ is attached to a carbon atom bonded to X, thenW′ is —C(O)NR¹³R¹⁴ or —C(O)OR¹²;

A is selected from —S(O)₂CH₂C(O)N(R³)—, —N{C(O)R⁵}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃ alkyl}—, —N{S(O)₂NR^(6a)R^(6b)}—,—S(O)₂N(R^(7a))—, and —OC(O)N(R^(7b))—;

R³ and R⁴ are independently C₁₋₄alkyl;

R⁵ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, C₄₋₆cycloalkyl, orpyrimidin-4-yl;

R^(6a) and R^(6b) are independently hydrogen, C₅₋₆cycloalkyl, orC₁₋₄alkyl, wherein C₁₋₄alkyl is optionally substituted with hydroxy,C₁₋₃alkoxy, or cyano;

R^(7a) and R^(7b) are independently hydrogen or C₁₋₄alkyl;

R⁸ is hydrogen or C₁₋₄alkyl;

R⁹ is hydrogen, furanyl, tetrahydrofuranyl, pyridinyl, or C₁₋₄alkyl;

R¹⁰ is C₁₋₄alkyl, optionally substituted with S(O)₂C₁₋₃alkyl, or withfrom 1 to 3 halo;

R¹¹ is —NR¹³R¹⁴, or C₁₋₄alkyl;

R¹² is C₁₋₄alkyl;

R¹³, R¹⁴ and R¹⁵ are independently hydrogen or C₁₋₄alkyl;

R¹⁶ is —(CH₂)_(r)—R¹⁷, wherein r is 0, 1, 2, or 3;

R¹⁷ is hydrogen, hydroxy, cyano, C₁₋₃alkyl, C₁₋₃alkoxy, —C(O)NR¹³R¹⁴,—CF₃, pyrrolyl, pyrrolidinyl, pyridinyl, tetrahydrofuranyl,—N(R⁸)C(O)OR¹², —OC(O)NR¹³R¹⁴, —N(R⁸)S(O)₂CH₃, —S(O)₂NR¹³R¹⁴, or2-oxoimidazolidin-1-yl, wherein C₁₋₃alkoxy is optionally substitutedwith hydroxy; provided that when r is 0, R¹⁷ is selected from hydrogen,C₁₋₃alkyl, and pyridinyl; and when r is 1, R¹⁷ is hydrogen or R¹⁷ formsa carbon-carbon bond with the —(CH₂)_(r)— carbon atom;

R¹⁸ is C₁₋₃alkyl optionally substituted with hydroxy;

or a pharmaceutically-acceptable salt or solvate or stereoisomerthereof.

The invention also provides a pharmaceutical composition comprising acompound of the invention and a pharmaceutically-acceptable carrier.

The invention also provides a method of treating a disease or conditionassociated with 5-HT₄ receptor activity, e.g. a disorder of reducedmotility of the gastrointestinal tract, the method comprisingadministering to the mammal, a therapeutically effective amount of acompound of the invention.

Further, the invention provides a method of treating a disease orcondition associated with 5-HT₄ receptor activity in a mammal, themethod comprising administering to the mammal, a therapeuticallyeffective amount of a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of the invention.

The compounds of the invention can also be used as research tools, i.e.to study biological systems or samples, or for studying the activity ofother chemical compounds. Accordingly, in another of its method aspects,the invention provides a method of using a compound of formula (I), or apharmaceutically acceptable salt or solvate or stereoisomer thereof, asa research tool for studying a biological system or sample or fordiscovering new 5-HT₄ receptor agonists, the method comprisingcontacting a biological system or sample with a compound of theinvention and determining the effects caused by the compound on thebiological system or sample.

In separate and distinct aspects, the invention also provides syntheticprocesses and intermediates described herein, which are useful forpreparing compounds of the invention.

The invention also provides a compound of the invention as describedherein for use in medical therapy, as well as the use of a compound ofthe invention in the manufacture of a formulation or medicament fortreating a disease or condition associated with 5-HT₄ receptor activity,e.g. a disorder of reduced motility of the gastrointestinal tract, in amammal.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides novel quinolinone-carboxamide 5-HT₄ receptoragonists of formula (I), or pharmaceutically-acceptable salts orsolvates or stereoisomers thereof. These compounds may contain one ormore chiral centers and, when such a chiral center or centers arepresent, this invention is directed to racemic mixtures, purestereoisomers; and stereoisomer-enriched mixtures of such isomers,unless otherwise indicated. When a particular stereoisomer is shown, itwill be understood by those skilled in the art, that minor amounts ofother stereoisomers may be present in the compositions of the inventionunless otherwise indicated, provided that any utility of the compositionas a whole is not eliminated by the presence of such other isomers.

The compounds of this invention also contain several basic groups (e.g.,amino groups) and therefore, the compounds of formula (I) and itsintermediates can exist in various salt forms. All such salt forms areincluded within the scope of this invention. Also, included within thescope of this invention are pharmaceutically-acceptable solvates of thecompounds of formula (I) or the salts thereof.

REPRESENTATIVE EMBODIMENTS

The following substituents and values are intended to providerepresentative examples and embodiments of various aspects of thisinvention. These representative values are intended to further definesuch aspects and embodiments and are not intended to exclude otherembodiments or limit the scope of this invention. In this regard, therepresentation herein that a particular value or substituent ispreferred is not intended in any way to exclude other values orsubstituents from this invention unless specifically indicated.

In a specific aspect, R¹ is hydrogen or halo.

In another specific aspect, R¹ is hydrogen, bromo, fluoro, or methyl. Inanother specific aspect, R¹ is hydrogen.

In a specific aspect, R² is n-propyl, isopropyl, n-butyl, sec-butyl,tert-butyl, cyclobutyl or cyclopentyl.

In another specific aspect, R² is C₃₋₄alkyl.

In still another specific aspect, R² is isopropyl.

In a specific aspect, X is carbon. In specific aspects, X is carbon andQ is -A(CH₂)₂N(R⁴)—; or X is carbon and Q is -A-. When X is carbon,representative Q groups include —N{C(O)R⁵}—, —N{C(O)NR^(6a)R^(6b)}—,—N{S(O)₂C₁₋₃alkyl}—, and —S(O)₂N(R^(7a))—, such as —N{C(O)C₁₋₃alkoxy}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}—, and —S(O)₂N(R^(7a))—.

In another specific aspect, X is nitrogen. In specific aspects, X isnitrogen and Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,-A(CH₂)₂—,

or X is nitrogen and Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,and -A(CH₂)₂—.

In another aspect, when X is nitrogen, Q is selected from —OC(O)—,—S(O)₂—, —S(O)₂(CH₂)₂—, —S(O)₂N(R^(7a))(CH₂)₂—, —N{C(O)R⁵}(CH₂)₂—, and—N{S(O)₂C₁₋₃alkyl}(CH₂)₂—. When X is nitrogen, representative Q moietiesinclude —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, —S(O)₂N(CH₃)(CH₂)₂—,—N{C(O)CH₃}(CH₂)₂—, —N{C(O)OCH₃}(CH₂)₂— and —N{S(O)₂CH₃}(CH₂)₂—.

In yet another aspect, when X is nitrogen, Q is selected from —OC(O)—,—S(O)₂—, —S(O)₂(CH₂)₂—, —S(O)₂N(R^(7a))(CH₂)₂—,—N{C(O)C₁₋₃alkoxy}(CH₂)₂—, and —N{S(O)₂C₁₋₃alkyl}(CH₂)₂—.

In a specific aspect, A is selected from —N{C(O)R⁵}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}—, and —S(O)₂N(R^(7a))—.

In another specific aspect, A is selected from —N{C(O)C₁₋₃alkyl}—,—N{C(O)C₁₋₃alkoxy}-, —N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}-, and—S(O)₂N(R^(7a))—.

In still another specific aspect, A is selected from —N{C(O)CH₃}—,—N{C(O)OCH₃}—; —N{C(O)NH₂}—, —N{C(O)NHCH₃}—, —N{C(O)N(CH₃)₂}—,—N{S(O)₂CH₃}—, and —S(O)₂N(CH₃)—.

In a specific aspect, G is W and c is 0, wherein W is as defined informula (I). In another specific aspect, G is W, c is 0, b is 1, and Xis carbon.

In a specific aspect, W is selected from —N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—,—N{C(O)NR¹³R¹⁴}—, —N{R¹⁶}—, and —S(O)₂—, such as—N{C(O)-tetrahydrofuran-2-yl}-, —N{C(O)CH₃}—, —N{C(O)CH₂CH₃}—,—N{S(O)₂CH₃}—, —N{S(O)₂CH₂CH₃}—, —N{C(O)NH₂}—, —N{C(O)NHCH₃}—,—N{C(O)N(CH₃)₂}—, —N{CH₃}—, —N{(CH₂)₂CN}—, —N{(CH₂)₂CH₃}—, and —S(O)₂—.In another specific aspect, W is selected from —S(O)₂—, —N{C(O)R⁹}—,—N{S(O)₂R¹⁰}—, —N{C(O)NR¹³R¹⁴}—, and —N{R¹⁶}—.

Alternatively, in another specific aspect, G is carbon, c is 1, and Y isa moiety of formula (b). In another specific aspect, G is carbon, c is1, X is nitrogen, and Y is a moiety of formula (b).

In a specific aspect, W′ is selected from —N(R⁸)C(O)R⁹, —N(R⁸)S(O)₂R¹⁰,—S(R¹¹)(O)₂, —N(R⁸)C(O)NR¹³R¹⁴, —OR¹⁵, and —N(R⁸)R¹⁶, such as—N(CH₃)C(O)CH₃, —NHC(O)CH₃, —N(CH₃)C(O)H, —N(CH₃)C(O)CH₂CH₃,—N(CH₃)S(O)₂CH₃, —N(CH₃)S(O)₂CH₂CH₃, —S(O)₂CH₃, —N(CH₃)C(O)NH₂,—N(CH₃)C(O)NHCH₃, —N(CH₃)C(O)N(CH₃)₂, —OH, —OCH₃, —N(CH₃)₂,—N(CH₃)(CH₂)₂CN, and —N(CH₃)(CH₂)₂CH₃. In another specific aspect, W′ isselected from —OR¹⁵ and —N(R⁸)R¹⁶.

In a specific aspect, R³ and R⁴ are independently C₁₋₃alkyl, such asmethyl or ethyl. In another specific aspect, R³ and R⁴ are methyl.

In a specific aspect, R⁵ is hydrogen, C₁₋₃alkyl, or C₁₋₃alkoxy, such ashydrogen, methyl, or methoxy. In other specific aspects, R⁵ isC₁₋₃alkyl, such as methyl; or R⁵ is C₁₋₃alkoxy, such as methoxy.

In a specific aspect, R^(6a) and R^(6b) are independently hydrogen orC₁₋₄alkyl, for instance, R^(6a) and R^(6b) are independently hydrogen ormethyl.

In a specific aspect, R^(7a) and R^(7b) are independently hydrogen orC₁₋₄alkyl, such as hydrogen or methyl. In another specific aspect,R^(7a) and R^(7b) are methyl.

In a specific aspect, R⁸ is hydrogen or C₁₋₃alkyl, such as hydrogen,methyl, or ethyl. In a specific aspect, R⁸ is hydrogen. In anotherspecific aspect, R⁸ is methyl.

In a specific aspect, R⁹ is tetrahydrofuranyl, methyl, or ethyl.

In specific aspects, R¹⁰, R¹¹, and R¹² are independently methyl orethyl; or R¹⁰, R¹¹, and R¹² are methyl.

In a specific aspect, R¹³, and R¹⁴ are independently hydrogen, methyl orethyl. In another specific aspect, R¹³, and R¹⁴ are independentlyhydrogen or methyl.

In a specific aspect, R¹⁵ is hydrogen or C₁₋₃alkyl, such as hydrogen ormethyl. In a specific aspect, R¹⁵ is hydrogen. In another specificaspect, R¹⁵ is methyl.

In a specific aspect, R¹⁶ is —(CH₂)_(r)—R¹⁷, wherein r is 0, or 1, or 2.In other specific aspects, r is 0; r is 1; or r is 2.

In a specific aspect, R¹⁷ is selected from hydroxy, cyano, C₁₋₃alkyl,and C₁₋₃alkoxy. In other specific aspects, R¹⁷ is selected from hydroxy,cyano, methyl, ethyl, propyl, methoxy, and ethoxy; or R¹⁷ is selectedfrom cyano, methyl, ethyl, and propyl.

In a specific aspect, R¹⁸ is methyl or ethyl, wherein methyl or ethyl isoptionally substituted with hydroxy.

In specific aspects, a is 0; or a is 1.

In specific aspects, b is 1 or 2; or b is 1; or b is 2.

In a specific aspect, b is 1 or 2, X is carbon, G is W, c is 0, and W is—S(O)₂—.

In another specific aspect, b is 2, G is W and c is 0, and W is selectedfrom —S(O)₂—, —N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—, —N{C(O)NR¹³R¹⁴}—, and—N{R¹⁶}—.

In specific aspects, c is 0; or c is 1.

In a specific aspect, d is 0.

In specific aspects, e is 0; or e is 1.

The invention further provides a compound of formula (I), wherein R¹ ishydrogen or halo, R² is C₃₋₄alkyl, and d is 0.

The invention further provides a compound of formula (I), wherein X iscarbon and Q is -A-; or X is nitrogen and Q is selected from —OC(O)—,—S(O)₂—, —S(O)₂(CH₂)₂—, and -A(CH₂)₂—.

The invention further provides a compound of formula (I), wherein X iscarbon and Q is selected from —N{C(O)R⁵}—, —N{C(O)NR^(6a)R^(6b)}—,—N{S(O)₂C₁₋₃alkyl}-, and —S(O)₂N(R^(7a))—; or X is nitrogen and Q isselected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, —S(O)₂N(R^(7a))(CH₂)₂—,—N{C(O)R⁵}(CH₂)₂—, and —N{S(O)₂C₁₋₃alkyl}(CH₂)₂—.

The invention further provides a compound of formula (I), wherein G is Wand c is 0, wherein W is selected from —N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—,—N{C(O)NR¹³R¹⁴}—, —N{R¹⁶}—, and —S(O)₂—; or G is carbon, c is 1, and Yis a moiety of formula (b), wherein W′ is selected from —N(R⁸)C(O)R⁹,—N(R⁸)S(O)₂R¹⁰, —S(R¹¹)(O)₂, —N(R⁸)C(O)NR¹³R¹⁴, —OR¹⁵, and —N(R⁸)R¹⁶.

Additionally, the invention provides a compound of formula (I), whereinZ is:

(i) a moiety of formula (c):

wherein:

X is carbon and Q is -A-;

or X is nitrogen and Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,and -A(CH₂)₂—;

b is 1, X is carbon, and W is —S(O)₂—;

or b is 2, X is carbon or nitrogen, and W is selected from —S(O)₂—,—N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—, —N{C(O)NR¹³R¹⁴}—, and —N{R¹⁶}—; or

(ii) a moiety of formula (d):

wherein:

Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, and -A(CH₂)₂—;

b is 1 or 2; and

W′ is selected from —N(R⁸)C(O)R⁹, —N(R⁸)S(O)₂R¹⁰, —S(R¹¹)(O)₂,—N(R⁸)C(O)NR¹³R¹⁴, —OR¹⁵, and —N(R⁸)R⁶; and

R⁸ is hydrogen, methyl, or ethyl;

R⁹ is tetrahydrofuranyl, methyl, or ethyl;

R¹⁰ is methyl or ethyl;

R¹¹ is methyl or ethyl;

R¹³ and R¹⁴ are independently hydrogen, methyl or ethyl;

R¹⁵ is hydrogen or methyl;

R¹⁶ is —(CH₂)_(r)—R¹⁷, wherein r is 0, 1, or 2; and

R¹⁷ is selected from hydroxy, cyano, C₁₋₃alkyl, and C₁₋₃alkoxy.

The invention also provides a compound of formula (I):

wherein:

R¹ is hydrogen, halo, or C₁₋₄alkyl;

R² is C₃₋₄alkyl or C₃₋₆cycloalkyl;

a is 0 or 1;

Z is a moiety of formula (c):

wherein:

X is carbon and Q is -A-;

or X is nitrogen and Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,and -A(CH₂)₂—;

b is 1, X is carbon, and W is —S(O)₂—;

or b is 2, X is carbon or nitrogen, and W is selected from —S(O)₂—,—N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—, —N{C(O)NR¹³R¹⁴}—, and —N{R¹⁶}—; or

Z is a moiety of formula (d):

wherein:

Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, and -A(CH₂)₂—;

b is 1 or 2; and

W′ is selected from —N(R⁸)C(O)R⁹, —N(R⁸)S(O)₂R¹⁰, —S(R¹¹)(O)₂,—N(R⁸)C(O)NR¹³R¹⁴, —OR¹⁵, and —N(R⁸)R¹⁶; provided that when W′ isattached to a carbon atom bonded to the nitrogen atom of the ring, thenW′ is —C(O)NR¹³R¹⁴; and

A is selected from —S(O)₂CH₂C(O)N(R³)—, —N{C(O)R⁵}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}-, —N{S(O)₂NR^(6a)R^(6b)}—,—S(O)₂N(R^(7a))—, and —OC(O)N(R^(7b))—;

R³ is C₁₋₄alkyl;

R⁵ is hydrogen, C₁₋₃alkyl, or C₁₋₃alkoxy;

R^(6a) and R^(6b) are independently hydrogen or C₁₋₄alkyl;

R^(7a) and R^(7b) are independently hydrogen or C₁₋₄alkyl;

R⁸ is hydrogen, methyl, or ethyl;

R⁹ is tetrahydrofuranyl, methyl, or ethyl;

R¹⁰ is methyl or ethyl;

R¹¹ is methyl or ethyl;

R¹³ and R¹⁴ are independently hydrogen, methyl or ethyl;

R¹⁵ is hydrogen or methyl;

R¹⁶ is —(CH₂)_(r)—R⁷, wherein r is 0, 1, or 2; and R¹⁷ is selected fromhydroxy, cyano, C₁₋₃alkyl, and C₁₋₃alkoxy; provided that when r is 0,R¹⁷ is selected from C₁₋₃alkyl; and when r is 1, R¹⁷ is cyano orC₁₋₃alkyl;

or a pharmaceutically-acceptable salt or solvate or stereoisomerthereof.

In a specific aspect, Z is a moiety of formula (c).

In specific aspects, Z is a moiety of formula (c), wherein X isnitrogen, b is 2, and Q is selected from —OC(O)—, —S(O)₂—,—S(O)₂(CH₂)₂—, —S(O)₂N(R^(7a))(CH₂)₂—, —N{C(O)C₁₋₃alkoxy}(CH₂)₂— and—N{S(O)₂C₁₋₃alkyl}(CH₂)₂—; or Z is a moiety of formula (c), wherein X isnitrogen, b is 2, and Q is selected from —OC(O)—, —S(O)₂—,—S(O)₂(CH₂)₂—, and —N{C(O)C₁₋₃alkoxy}(CH₂)₂—.

In other specific aspects, Z is a moiety of formula (c), wherein X isnitrogen, b is 2, and Q is —OC(O)—; Z is a moiety of formula (c),wherein X is nitrogen, b is 2, and Q is —S(O)₂—; or Z is a moiety offormula (c), wherein X is nitrogen, b is 2, and Q is —S(O)₂(CH₂)₂—.

In another aspect, Z is a moiety of formula (c), wherein X is nitrogen,Q is as defined herein, b is 2, and W is selected from —N{C(O)R⁹}—, and—N{S(O)₂R¹¹}—.

In another aspect, Z is a moiety of formula (c), wherein X is carbon andQ is selected from —N{C(O)C₁₋₃alkoxy}-, —N{C(O)NR^(6a)R^(6b)}—,—N{S(O)₂C₁₋₃alkyl}-, and —S(O)₂N(R^(7a))—.

In other specific aspects, Z is a moiety of formula (c), wherein X iscarbon and Q is —N{C(O)C₁₋₃alkoxy}-; Z is a moiety of formula (c),wherein X is carbon and Q is —N{C(O)NR^(6a)R^(6b)}—; or Z is a moiety offormula (c), wherein X is carbon and Q is —N{S(O)₂R¹⁰}—.

In still another specific aspect, Z is a moiety of formula (c), whereinX is carbon, Q is as defined herein, and W is —S(O)₂—.

In a specific aspect, Z is a moiety of formula (d).

In yet another aspect, Z is a moiety of formula (d), wherein Q isselected from —OC(O)— and —S(O)₂—.

In another specific aspect, Z is a moiety of formula (d), wherein W′ isselected from —OR¹⁵ and —N(R⁸)R¹⁶.

Included within the invention are the compounds listed in Tables 1 to 5herein.

The chemical naming conventions used herein are illustrated for thecompound of Example 28:

which is designated 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid[(1R,3R,5S)-8-(2-{[2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]methanesulfonylamino}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]-amide,according to the AutoNom software, provided by MDL Information Systems,GmbH (Frankfurt, Germany). The designation (1S,3R,5R) describes therelative orientation of the bonds associated with the bicyclic ringsystem that are depicted as solid and dashed wedges. The compound isalternatively denoted asN-[(3-endo)-8-(2-{[2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]methanesulfonylamino}ethyl)-8-azabicyclo-[3.2.1]oct-3-yl]-1-(1-methylethyl)-2-oxo-1,2-dihydro-3-quinolinecarboxamide.In all of the compounds of the invention listed by name below, thequinolinone-carboxamide is endo to the azabicyclooctyl group.

Particular mention may be made of the following compounds:

-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(4-methanesulfonylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(3-dimethylaminopyrrolidine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(4-methylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[methanesulfonyl-(1-propylpiperidin-4-yl)amino]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)-amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    [(1S,3R,5R)-8-(3-{[1-(2-methoxyethyl)piperidin-4-yl]methylsulfamoyl}propyl)-8-azabicyclo[3.2.1]oct-3-yl]amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[(1-methanesulfonylpiperidin-4-yl)methylsulfamoyl]propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    [(1S,3R,5R)-8-(3-{[1-(2-cyanoethyl)piperidin-4-yl]methylsulfamoyl}propyl)-8-azabicyclo[3.2.1]oct-3-yl]amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)methanesulfonylamino]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3,3-dimethylureido]ethyl}-8-azabicyclo[3.2.1]-oct-3-yl)amide;-   (1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamic    acid methyl ester;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    [(1S,3R,5R)-8-(2-{[2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]methanesulfonylamino}ethyl)-8-azabicyclo-[3.2.1]oct-3-yl]amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[2-(4-methanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    [(1S,3R,5R)-8-(2-{2-[4-(tetrahydrofuran-2-carbonyl)piperazin-1-yl]ethanesulfonyl}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[2-(4-ethanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   (1,1-dioxo-hexahydro-1λ⁶-thiopyran-4-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamic    acid methyl ester;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3-methylureido]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;-   (2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)-amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-[2-(4-methanesulfonylpiperazin-1-ylethyl]-carbamic    acid methyl ester;-   [2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-ethyl)-carbamic    acid methyl ester;-   [2-(4-acetyl-piperazin-1-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamic    acid methyl ester;-   [2-(1,1-dioxo-1λ⁶-thiomorpholin-4-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamic    acid methyl ester;

(1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamicacid methyl ester;

-   ((S)-1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamic    acid methyl ester;-   1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(methyl-{2-[4-(tetrahydrofuran-2-carbonyl)piperazin-1-yl]ethyl}sulfamoyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    ((1S,3R,5R)-8-{3-[4-(tetrahydrofuran-2-carbonyl)piperazine-1-sulfonyl]propyl}-8-aza-bicyclo-[3.2.1]oct-3-yl)amide;-   1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid    {(1S,3R,5R)-8-[3-(4-acetylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;-   4-methanesulfonyl-piperazine-1-carboxylic acid    3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl    ester;-   4-(tetrahydrofuran-2-carbonyl)piperazine-1-carboxylic acid    3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-aza-bicyclo[3.2.1]oct-8-yl}propyl    ester;-   4-acetyl-piperazine-1-carboxylic acid    3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl    ester; and-   4-hydroxypiperidine-1-carboxylic acid    3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-propyl    ester.

DEFINITIONS

When describing the compounds, compositions and methods of theinvention, the following terms have the following meanings, unlessotherwise indicated.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched or combinations thereof. Examples ofparticular values for a C₁₋₄alkyl group include, by way of example,methyl, ethyl, n-propyl (n-Pr), isopropyl (i-Pr), n-butyl (n-Bu),sec-butyl, isobutyl, and tert-butyl.

The term “alkylene” means a divalent saturated hydrocarbon group whichmay be linear or branched or combinations thereof. Examples ofparticular values for a C₂₋₅alkylene include ethylene, propylene,isopropylene, butylene, and pentylene, and the like.

The term “alkoxy” means a monovalent group —O-alkyl, where alkyl isdefined as above. Representative alkoxy groups include, by way ofexample, methoxy, ethoxy, propoxy, butoxy, and the like.

The term “cycloalkyl” means a monovalent saturated carbocyclic groupwhich may be monocyclic or multicyclic. Unless otherwise defined, suchcycloalkyl groups typically contain from 3 to 10 carbon atoms.Representative C₃₋₆cycloalkyl groups include, by way of example,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term “halo” means a fluoro, chloro, bromo or iodo.

The term “compound” means a compound that was synthetically prepared orprepared in any other way, such as by metabolism.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need of treatment.

The term “treatment” as used herein means the treatment of a disease,disorder, or medical condition in a patient, such as a mammal(particularly a human) which includes:

-   -   (a) preventing the disease, disorder, or medical condition from        occurring, i.e., prophylactic treatment of a patient;    -   (b) ameliorating the disease, disorder, or medical condition,        i.e., eliminating or causing regression of the disease,        disorder, or medical condition in a patient;    -   (c) suppressing the disease, disorder, or medical condition,        i.e., slowing or arresting the development of the disease,        disorder, or medical condition in a patient; or    -   (d) alleviating the symptoms of the disease, disorder, or        medical condition in a patient.

The term “pharmaceutically-acceptable salt” means a salt prepared froman acid or base which is acceptable for administration to a patient,such as a mammal. Such salts can be derived frompharmaceutically-acceptable inorganic or organic acids and frompharmaceutically-acceptable bases. Typically,pharmaceutically-acceptable salts of compounds of the present inventionare prepared from acids.

Salts derived from pharmaceutically-acceptable acids include, but arenot limited to, acetic, benzenesulfonic, benzoic, camphorsulfonic,citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic,hydrochloric, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pantothenic, phosphoric, succinic, sulfuric, tartaric,p-toluenesulfonic, xinafoic (1-hydroxy-2-naphthoic acid),napthalene-1,5-disulfonic acid and the like.

The term “solvate” means a complex or aggregate formed by one or moremolecules of a solute, i.e. a compound of the invention or apharmaceutically-acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include by way of example, water, methanol, ethanol,isopropanol, acetic acid, and the like. When the solvent is water, thesolvate formed is a hydrate.

It will be appreciated that the term “or a pharmaceutically-acceptablesalt or solvate of stereoisomer thereof” is intended to include allpermutations of salts, solvates and stereoisomers, such as a solvate ofa pharmaceutically-acceptable salt of a stereoisomer of a compound offormula (I).

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include halo, such as chloro,bromo and iodo groups; sulfonic ester groups, such as mesylate,tosylate, brosylate, nosylate and the like; and acyloxy groups, such asacetoxy, trifluoroacetoxy and the like.

The term “amino-protecting group” means a protecting group suitable forpreventing undesired reactions at an amino nitrogen. Representativeamino-protecting groups include, but are not limited to, formyl; acylgroups, for example alkanoyl groups, such as acetyl; alkoxycarbonylgroups, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl groups,such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc);arylmethyl groups, such as benzyl (Bn), trityl (Tr), and1,1-di-(4′-methoxyphenyl)methyl; silyl groups, such as trimethylsilyl(TMS) and tert-butyldimethylsilyl (TBDMS); and the like.

The term “hydroxy protecting group” means a protecting group suitablefor preventing undesired reactions of a hydroxyl group. The term“hydroxyl-protecting group” means a protecting group suitable forpreventing undesirable reactions at a hydroxyl group. Representativehydroxyl-protecting groups include, but are not limited to, silyl groupsincluding tri(1-6C)alkylsilyl groups, such as trimethylsilyl (TMS),triethylsilyl (TES), tert-butyldimethylsilyl (TBS) and the like; esters(acyl groups) including (1-6C)alkanoyl groups, such as formyl, acetyland the like; arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl(PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM) and thelike. Additionally, two hydroxyl groups can also be protected as analkylidene group, such as prop-2-ylidine, formed, for example, byreaction with a ketone, such as acetone.

General Synthetic Procedures

Compounds of the invention can be prepared from readily availablestarting materials using the following general methods and procedures.Although a particular aspect of the present invention is illustrated inthe schemes below, those skilled in the art will recognize that allaspects of the present invention can be prepared using the methodsdescribed herein or by using other methods, reagents and startingmaterials known to those skilled in the art. It will also be appreciatedthat where typical or preferred process conditions (i.e., reactiontemperatures, times, mole ratios of reactants, solvents, pressures,etc.) are given, other process conditions can also be used unlessotherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group, as well assuitable conditions for protection and deprotection, are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and G. M. Wuts, ProtectingGroups in Organic Synthesis, Third Edition, Wiley, New York, 1999, andreferences cited therein.

The substituents and variables shown in the following schemes have thedefinitions provided herein unless otherwise indicated.

In one method of synthesis, compounds of formula (I) are prepared asillustrated in Scheme A:

As shown in Scheme A, a compound of formula (III) is reacted with acompound of formula (IV) wherein L¹ is a leaving group, such as halo,for example, chloro, or a sulfonic ester group, such as mesylate,tosylate, brosylate, nosylate and the like, to provide a compound offormula (I) or a salt or solvate or stereoisomer thereof.

When L¹ is a halo leaving group, such as chloro, the reaction istypically conducted by contacting a compound of formula (III) withbetween about 1 and about 4 equivalents of a compound of formula (IV) inan inert diluent, such as N,N-dimethylformamide (DMF), in the presenceof an excess of a base, for example between about 3 and about 6equivalents of base, such as N,N-diisopropylethylamine or1,8-diazabicyclo-[5.4.0]undec-7-ene (DBU), in the presence of acatalyst, such as sodium iodide. Suitable inert diluents also includeDMF, dichloromethane, trichloromethane, 1,1,2,2-tetrachloroethane,tetrahydrofuran, methanol, ethanol, and the like. Suitable catalystsinclude, for example, sodium iodide, potassium iodide, andtetrabutylammonium iodide. The reaction is typically conducted at atemperature in the range of about 15° C. to about 90° C. for about 4hours to about 48 hours, or until the reaction is substantiallycomplete.

The product of formula (I) is isolated and purified by conventionalprocedures. For example, the product can be concentrated to drynessunder reduced pressure, taken up in an aqueous weak acid solution andpurified by HPLC chromatography.

Alternatively, compounds of formula (I), wherein X is carbon and Q isselected from -A(CH₂)₂N(R⁴)— and —S(O)₂(CH₂)₂N(R⁴)—; or X is nitrogenand Q is selected from —S(O)₂(CH₂)₂— and -A(CH₂)₂—; can be prepared asillustrated in Scheme B shown below, to provide a compound of formula(I-a).

As shown in Scheme B, a compound of formula (V), wherein Q¹ is selectedfrom —S(O)₂— and -A-, is reacted with H-D, an amine compound of formula(VI), wherein D is selected from a moiety of formula (D1):

and a moiety of formula (D2):

to provide a compound of formula (I-a) or a salt or solvate orstereoisomer thereof.

It will be understood that while intermediate compound (V) is shown inthe form of an aldehyde hydrate, intermediate (V) can equivalently bedepicted in the form of an aldehyde.

In scheme B, intermediate compound (V) is reductively coupled with anamine of formula (VI) to provide a compound of formula (I-a). Typically,a solution is prepared of between about 1 and about 3 equivalents of theamine of formula (VI) and a reducing agent in an inert diluent in thepresence of a base, such as, for example, N,N-diisopropylethylamine or1,8-diazabicyclo-[5.4.0]undec-7-ene (DBU). Suitable reducing agents, forexample, include hydrogen in the presence of a Group VIII metalcatalyst, such as palladium on charcoal, or a borohydride, such assodium triacetoxyborohydride, sodium cyanoborohydride, lithiumcyanoborohydride, and the like. Suitable inert diluents includeacetonitrile, halogenated hydrocarbons, such as dichloromethane (DCM)and dichloroethane, alcohols, such as methanol, ethanol, and isopropylalcohol, or mixtures thereof.

Intermediate (V) is added slowly to the amine mixture. Generally, thisreaction is conducted at a temperature ranging from about 0° C. to about50° C. for a period of about 10 minutes to about 12 hours or until thereaction is substantially complete. The reaction product is thenisolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

Alternatively, a compound of formula (I-a), wherein Q¹ is —S(O)₂—, canbe prepared by reacting an intermediate compound (V-a):

with a compound of formula (VI), to provide a compound of formula (I-a),wherein Q¹ is —S(O)₂—, or a salt or solvate or stereoisomer thereof.This reaction is typically conducted either in the presence of a base,such as N,N′-diisopropylethylamine or inorganic bases, such as sodiumhydroxide, and potassium hydroxide when the reacting amines are given insalt form, or in the absence of a base when the reacting amines aregiven in neutral form. Generally, this reaction is conducted in an inertdiluent, such as dichloromethane, methanol, ethanol, DMF, or water, at atemperature ranging from about 0° C. to about 100° C. until the reactionis substantially complete. The reaction product is then isolated usingconventional procedures, such as extraction, recrystallization,chromatography and the like.

A compound of formula (I-a) can also be prepared by reacting anintermediate compound (V-b):

in which L² is a leaving group, with a compound of formula (VI), toprovide a compound of formula (I-a). Typical conditions for thiscoupling reaction are described in Scheme A.

Alternatively, compounds of formula (I), wherein X is carbon and Q isselected from -A¹- and -A¹(CH₂)₂N(R⁴)—; or X is nitrogen and Q isselected from -A¹(CH₂)₂—, wherein A¹- is selected from —N{C(O)R⁵}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}-, and—N{S(O)₂NR^(6a)R^(6b)}—; can be prepared as illustrated in Scheme Cshown below:

As shown in Scheme C, a compound of formula (VII), wherein E is selectedfrom a moiety of formula (E1):

and a moiety of formula —CH₂CH₂-D, wherein D is selected from a moietyof formula (D1) and a moiety of formula (D2); is reacted with a compoundof formula (VIII), wherein L³-R^(a) is C₁₋₄alkylisocyanate, or L³ is aleaving group, such as halo, p-nitrophenol, or a sulfonic ester group,and R^(a) is —C(O)R⁵, C(O)NR^(6a)R^(6b), —S(O)₂C₁₋₃alkyl, or—S(O)₂NR^(6a)R^(6b); to provide a compound of formula (I-b) or a salt orsolvate or stereoisomer thereof.

Typically, compound (VII) is contacted with between about 1 and about 6equivalents of compound (VIII) in an inert diluent, such asdichloromethane, chloroform, N-methylpyrrolidinone, DMF, or the like, inthe presence of 2 to 3 equivalents of a base, such asN,N-diisopropylethylamine, triethylamine, potassium carbonate, sodiumhydroxide, and the like. The reaction is typically conducted at atemperature of between about 0° C. and about 120° C. for between about10 minutes and about 24 hours, or until the reaction is substantiallycomplete to provide a compound of formula (I-b).

A compound of formula (III) can be prepared as illustrated in Scheme D:

In Scheme D, a substituted quinolinone carboxylic acid (1) is reactedwith a protected aminotropane (2), wherein P¹ is an amino-protectinggroup, to provide a protected intermediate (III-p), which is thende-protected to provide a compound of formula (III).

A substituted quinolinone carboxylic acid (1) can be readily prepared byprocedures similar to those reported in the literature in Suzuki et al,Heterocycles, 2000, 53, 2471-2485 and described in the examples below.

A protected aminotropane (2) or aminoazabicyclooctane can be preparedfrom readily available starting materials. For example, when theprotecting group P¹ is Boc, the protected tropane can be prepared bycontacting 2,5-dimethoxy tetrahydrofuran with between about 1 and 2equivalents, preferably about 1.5 equivalents of benzyl amine and aslight excess, for example about 1.1 equivalents, of1,3-acetonedicarboxylic acid in an acidic aqueous solution in thepresence of a buffering agent such as sodium hydrogen phosphate. Thereaction mixture is heated to between about 60° C. and about 100° C. toensure decarboxylation of any carboxylated intermediates in the product,8-benzyl-8-azabicyclo[3.2.1]octan-3-one, commonly N-benzyltropanone.

The resulting N-benzyltropanone is typically reacted with a slightexcess of di-tert-butyl dicarbonate (commonly (Boc)₂O), for example,about 1.1 equivalents, under a hydrogen atmosphere in the presence of atransition metal catalyst to provide a Boc protected intermediate,3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester. Thereaction is typically conducted at ambient temperature for about 12 toabout 72 hours. Finally, 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester is contacted with a large excess, for example atleast about 25 equivalents, of ammonium formate in an inert diluent,such as methanol, in the presence of a transition metal catalyst toprovide intermediate (2) where P¹ is Boc, in the endo configuration withhigh stereospecificity, for example, endo to exo ratio of >99:1. Thereaction is typically conducted at ambient temperature for about 12 toabout 72 hours or until the reaction is substantially complete. It isadvantageous to add the ammonium formate reagent in portions. Forexample, 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butylester is contacted with an initial portion of ammonium formate of about15 to about 25 equivalents. After an interval of about 12 to about 36hours, an additional portion of about 5 to about 10 equivalents ofammonium formate is added. The subsequent addition can be repeated aftera similar interval. The product can be purified by conventionalprocedures, such as alkaline extraction.

Intermediate compound (III) can be prepared by coupling a substitutedquinolinone carboxylic acid (1), with a protected aminotropane (2) underconditions similar to those described in Scheme A for amide bondformation. The protecting group P¹ can be removed by standard proceduresto provide an intermediate compound (III). For example when theprotecting group is Boc, typically removal is by treatment with an acid,such as trifluoroacetic acid, providing the acid salt of theintermediate. The protecting group Cbz, for another example, isconveniently removed by hydrogenolysis over a suitable metal catalystsuch as palladium on carbon.

An intermediate compound of formula (IV) can be prepared as illustratedbelow in Scheme E:

As shown in Scheme E, a compound of formula (IV) can be prepared byreacting an amine, intermediate (4), (6), or (8), with intermediate (3),(5), or (7) respectively, containing L⁴, a leaving group, to provide acompound of formula (IV). Q³ and Q⁴ are defined below.

For example, a compound of formula (IV) wherein X is carbon and Q isselected from —S(O)₂CH₂C(O)N(R³)—, —S(O)₂(CH₂)₂N(R⁴)—, —S(O)₂N(R^(7a))—,—OC(O)N(R^(7b))—, and -A(CH₂)₂N(R⁴)—, can be prepared by Scheme E-(i),by reacting intermediate (3) wherein L¹ and L⁴ are leaving groups, andQ³ is selected from —S(O)₂CH₂C(O)—, —S(O)₂(CH₂)₂—, —S(O)₂—, —OC(O)—, and-A(CH₂)₂—, with intermediate (4) wherein R^(b) is selected from R³, R⁴,R^(7a), and R^(7b) as defined herein; to provide a compound of formula(IV).

Similarly, a compound of formula (IV) wherein X is nitrogen and Q isselected from —S(O)₂CH₂C(O)—, —SCH₂C(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,—OC(O)—, -A(CH₂)₂—,

can be prepared by Scheme E-(ii) by reacting intermediate (5) wherein L¹and L⁴ are leaving groups, and Q⁴ is selected from —S(O)₂CH₂C(O)—,—SCH₂C(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, —OC(O)—, -A(CH₂)₂—,

with intermediate (6), to provide a compound of formula (IV).

Similarly, a compound of formula (IV) where X is carbon and Q is -A¹-,wherein A¹ is selected from —N{C(O)R⁵}—, —N{C(O)NR^(6a)R^(6b)}—,—N{S(O)₂C₁₋₃alkyl}-, and —N{S(O)₂NR^(6a)R^(6b)}—; can be prepared byScheme E-(iii); by reacting intermediate (7) wherein L¹ and L⁴ areleaving groups, with intermediate compound (8), wherein R^(a) is—C(O)R⁵, —C(O)NR^(6a)R^(6b), —S(O)₂C₁₋₃alkyl, or —S(O)₂NR^(6a)R^(6b); toprovide a compound of formula (IV).

The reactions of Scheme E are typically conducted under the conditionsdescribed above for Scheme A, and are further illustrated in theExamples herein.

A compound of formula (V) can be prepared as illustrated in Scheme F:

wherein a dimethoxy acetal intermediate compound (9) (wherein Q¹ isselected from —S(O)₂— and -A-) is hydrolyzed in an aqueous solution of astrong acid, for example, 3N or 6N HCl, to provide a compound of formula(V). While intermediate compound (V) is shown in the form of an aldehydehydrate, it can equivalently be depicted in the form of an aldehyde.

An intermediate compound (9-a), representative of intermediate (9),wherein a is 0, and Q¹ is selected from —N{C(O)R⁵}—,—N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}-, and—N{S(O)₂NR^(6a)R^(6b)}—, can be prepared as illustrated in Scheme G:

As shown in Scheme G, intermediate (III) is reductively N-alkylated byreaction with dimethoxyacetaldehyde to provide an intermediate offormula (10). This reaction is typically conducted by contactingintermediate (III) with between about 1 and about 4 equivalents ofdimethoxyacetaldehyde in an inert diluent in the presence of a base,such as N,N′-diisopropylethylamine, and between about 1 and about 2equivalents of a reducing agent. The reaction is typically conducted atambient temperature for about 1 to about 2 hours, or until the reactionis substantially complete. Suitable inert diluents includedichloromethane, trichloromethane, 1,1,2,2-tetrachloroethane, and thelike. Typical reducing agents include sodium triacetoxyborohydride,sodium borohydride, and sodium cyanoborohydride. The product (10) isisolated by standard procedures.

Next, the dimethoxy intermediate (10) is hydrolyzed in an aqueoussolution of a strong acid, for example 3N or 6H HCl, to provide thedihydroxyethyl intermediate (11). The reaction is typically conducted ata temperature in the range of about 25° C. to about 100° C. for about 15minutes to about 2 hours, or until the reaction is substantiallycomplete.

Next, intermediate (11) is reductively coupled with aminoacetaldehydedimethyl acetal, to provide intermediate (12). Typically a solution isprepared of between about 1 and about 2 equivalents of theaminoacetaldehyde dimethyl acetal and a reducing agent in an inertdiluent. The intermediate (11) is added slowly to the amine mixture. Thereaction is typically conducted at ambient temperature for about 15minutes to about 2 hours, or until the reaction is substantiallycomplete.

Finally intermediate (12) is reacted with a compound of formula (VIII)to provide intermediate (9-a). Typical conditions for this reaction aredescribed in Scheme C herein.

A compound of formula (VII) can be prepared as shown in Scheme H:

by reductively coupling a dihydroxy acetal intermediate (11) withintermediate (13) or (14), to provide a compound of formula (VII). Forexample, a compound of formula (VII) wherein E is a moiety of formula(E1) can be prepared by reductively coupling intermediate (11) withintermediate (14). Whereas a compound of formula (VII) wherein E is amoiety of the formula —CH₂CH₂-D can be prepared by reductively couplingintermediate (11) with intermediate (13). Typical conditions for thesereactions are described above in Scheme B.

Compounds of formulae (VI) and (VIII), and intermediates (3), (4), (5),(6), (7), (8), (13), and (14) employed in the reactions described inthis application are available commercially or are readily prepared bystandard procedures from common starting materials.

Further details regarding specific reaction conditions and otherprocedures for preparing representative compounds of the invention orintermediates thereto are described in the examples below.

Accordingly, the invention provides a process for preparing a compoundof formula (I):

wherein R¹, R², a and Z are as defined herein for a compound of formula(I), or a pharmaceutically-acceptable salt or solvate or stereoisomerthereof, the process comprising reacting a compound of formula (III):

or a salt or stereoisomer thereof, with a compound of formula (IV):

wherein L¹ is a leaving group, to provide a compound of formula (I) or apharmaceutically-acceptable salt or solvate or stereoisomer thereof.

The invention further provides a process for preparing a compound offormula (I-a):

wherein:

Q¹ is selected from —S(O)₂—, and -A-; and

D is selected from a moiety of formula (D1):

a moiety of formula (D2):

wherein R¹, R², R⁴, R¹⁸, A, Y, G, a, b, c, and d are as defined hereinfor a compound of formula (I); or a pharmaceutically-acceptable salt orsolvate or stereoisomer thereof, the process comprising reacting acompound of formula (V):

with a compound of formula (VI):H-D  (VI)

to provide a compound of formula (I-a) or a pharmaceutically-acceptablesalt or solvate or stereoisomer thereof.

Accordingly, the invention further provides a compound of formula (I-a).

The invention also provides a process for preparing a compound offormula (I-b):

wherein R^(a) is —C(O)R⁵, —C(O)NR^(6a)R^(6b), —S(O)₂C₁₋₃alkyl, or—S(O)₂NR^(6a)R^(6b); and

E is selected from a moiety of formula (E1):

and a moiety of formula —CH₂CH₂-D, wherein D is selected from a moietyof formula (DI):

and a moiety of formula (D2):

wherein R¹, R², R⁴, R⁵, R^(6a), R^(6b), R¹⁸, Y, G, a, b, c, and d are asdefined herein for a compound of formula (I); or apharmaceutically-acceptable salt or solvate or stereoisomer thereof, theprocess comprising reacting a compound of formula (VI):

with a compound of formula (VIII):L³-R^(a)  (VIII)

wherein L³-R^(a) is C₁₋₄alkylisocyanate, or L³ is a leaving group, andR^(a) is —C(O)R⁵, C(O)NR^(6a)R^(6b), —S(O)₂C₁₋₃alkyl, or—S(O)₂NR^(6a)R^(6b); to provide a compound of formula (I-b) or apharmaceutically-acceptable salt, solvate, or stereoisomer thereof.

In addition, the invention provides a compound of formula (I-b).

The invention further provides the product of the processes describedherein.

Pharmaceutical Compositions

The quinolinone-carboxamide compounds of the invention are typicallyadministered to a patient in the form of a pharmaceutical composition.Such pharmaceutical compositions may be administered to the patient byany acceptable route of administration including, but not limited to,oral, rectal, vaginal, nasal, inhaled, topical (including transdermal)and parenteral modes of administration.

Accordingly, in one of its compositions aspects, the invention isdirected to a pharmaceutical composition comprising apharmaceutically-acceptable carrier or excipient and a therapeuticallyeffective amount of a compound of formula (I) or apharmaceutically-acceptable salt thereof. Optionally, suchpharmaceutical compositions may contain other therapeutic and/orformulating agents if desired.

The pharmaceutical compositions of the invention typically contain atherapeutically effective amount of a compound of the present inventionor a pharmaceutically-acceptable salt thereof. Typically, suchpharmaceutical compositions will contain from about 0.1 to about 95% byweight of the active agent; preferably, from about 5 to about 70% byweight; and more preferably from about 10 to about 60% by weight of theactive agent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of the invention. The choice of a particular carrier orexcipient, or combinations of carriers or excipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable pharmaceutical composition for a particular mode ofadministration is well within the scope of those skilled in thepharmaceutical arts. Additionally, the ingredients for such compositionsare commercially available from, for example, Sigma, P.O. Box 14508, St.Louis, Mo. 63178. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of Pharmacy, 20^(th) Edition, Lippincott Williams & White,Baltimore, Md. (2000); and H. C. Ansel et al., Pharmaceutical DosageForms and Drug Delivery Systems, 7^(th) Edition, Lippincott Williams &White, Baltimore, Md. (1999).

Representative examples of materials which can serve aspharmaceutically-acceptable carriers include, but are not limited to,the following: (1) sugars, such as lactose, glucose and sucrose; (2)starches, such as corn starch and potato starch; (3) cellulose, such asmicrocrystalline cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalcompositions.

The pharmaceutical compositions of the invention are typically preparedby thoroughly and intimately mixing or blending a compound of theinvention with a pharmaceutically-acceptable carrier and one or moreoptional ingredients. If necessary or desired, the resulting uniformlyblended mixture can then be shaped or loaded into tablets, capsules,pills and the like using conventional procedures and equipment.

The pharmaceutical compositions of the invention are preferably packagedin a unit dosage form. The term “unit dosage form” means a physicallydiscrete unit suitable for dosing a patient, i.e., each unit containinga predetermined quantity of active agent calculated to produce thedesired therapeutic effect either alone or in combination with one ormore additional units. For example, such unit dosage forms may becapsules, tablets, pills, and the like.

In a preferred embodiment, the pharmaceutical compositions of theinvention are suitable for oral administration. Suitable pharmaceuticalcompositions for oral administration may be in the form of capsules,tablets, pills, lozenges, cachets, dragees, powders, granules; or as asolution or a suspension in an aqueous or non-aqueous liquid; or as anoil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup;and the like; each containing a predetermined amount of a compound ofthe present invention as an active ingredient.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the pharmaceutical compositionsof the invention will typically comprise a compound of the presentinvention as the active ingredient and one or morepharmaceutically-acceptable carriers, such as sodium citrate ordicalcium phosphate. Optionally or alternatively, such solid dosageforms may also comprise: (1) fillers or extenders, such as starches,microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as carboxymethylcellulose, alginates,gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants,such as glycerol; (4) disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and/or sodium carbonate; (5) solution retarding agents, such asparaffin; (6) absorption accelerators, such as quaternary ammoniumcompounds; (7) wetting agents, such as cetyl alcohol and/or glycerolmonostearate; (8) absorbents, such as kaolin and/or bentonite clay; (9)lubricants, such as talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and/or mixtures thereof;(10) coloring agents; and (11) buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants can also be presentin the pharmaceutical compositions of the invention. Examples ofpharmaceutically-acceptable antioxidants include: (1) water-solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodiumbisulfate, sodium metabisulfate sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal-chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like. Coating agents fortablets, capsules, pills and like, include those used for entericcoatings, such as cellulose acetate phthalate (CAP), polyvinyl acetatephthalate (PVAP), hydroxypropyl methylcellulose phthalate, methacrylicacid-methacrylic acid ester copolymers, cellulose acetate trimellitate(CAT), carboxymethyl ethyl cellulose (CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), and the like.

If desired, the pharmaceutical compositions of the present invention mayalso be formulated to provide slow or controlled release of the activeingredient using, by way of example, hydroxypropyl methyl cellulose invarying proportions; or other polymer matrices, liposomes and/ormicrospheres.

In addition, the pharmaceutical compositions of the present inventionmay optionally contain opacifying agents and may be formulated so thatthey release the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.Examples of embedding compositions which can be used include polymericsubstances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically-acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Such liquid dosage formstypically comprise the active ingredient and an inert diluent, such as,for example, water or other solvents, solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (such as, for example, cottonseed, groundnut,corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Suspensions, in addition to the active ingredient, maycontain suspending agents such as, for example, ethoxylated isostearylalcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

Alternatively, the pharmaceutical compositions of the invention areformulated for administration by inhalation. Suitable pharmaceuticalcompositions for administration by inhalation will typically be in theform of an aerosol or a powder. Such compositions are generallyadministered using well-known delivery devices, such as a metered-doseinhaler, a dry powder inhaler, a nebulizer or a similar delivery device.

When administered by inhalation using a pressurized container, thepharmaceutical compositions of the invention will typically comprise theactive ingredient and a suitable propellant, such asdichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas.

Additionally, the pharmaceutical composition may be in the form of acapsule or cartridge (made, for example, from gelatin) comprising acompound of the invention and a powder suitable for use in a powderinhaler. Suitable powder bases include, by way of example, lactose orstarch.

The compounds of the invention can also be administered transdermallyusing known transdermal delivery systems and excipients. For example, acompound of the invention can be admixed with permeation enhancers, suchas propylene glycol, polyethylene glycol monolaurate,azacycloalkan-2-ones and the like, and incorporated into a patch orsimilar delivery system. Additional excipients including gelling agents,emulsifiers and buffers, may be used in such transdermal compositions ifdesired.

The following formulations illustrate representative pharmaceuticalcompositions of the present invention:

Formulation Example A

Hard gelatin capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 50 mg Lactose (spray-dried)200 mg Magnesium stearate 10 mg

Representative Procedure: The ingredients are thoroughly blended andthen loaded into a hard gelatin capsule (260 mg of composition percapsule).

Formulation Example B

Hard gelatin capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 20 mg Starch 89 mgMicrocrystalline cellulose 89 mg Magnesium stearate 2 mg

Representative Procedure: The ingredients are thoroughly blended andthen passed through a No. 45 mesh U.S. sieve and loaded into a hardgelatin capsule (200 mg of composition per capsule).

Formulation Example C

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 10 mg Polyoxyethylenesorbitan monooleate 50 mg Starch powder 250 mg

Representative Procedure: The ingredients are thoroughly blended andthen loaded into a gelatin capsule (310 mg of composition per capsule).

Formulation Example D

Tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 5 mg Starch 50 mgMicrocrystalline cellulose 35 mg Polyvinylpyrrolidone (10 wt. % inwater) 4 mg Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mgTalc 1 mg

Representative Procedure: The active ingredient, starch and celluloseare passed through a No. 45 mesh U.S. sieve and mixed thoroughly. Thesolution of polyvinylpyrrolidone is mixed with the resulting powders,and this mixture is then passed through a No. 14 mesh U.S. sieve. Thegranules so produced are dried at 50-60° C. and passed through a No. 18mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate andtalc (previously passed through a No. 60 mesh U.S. sieve) are then addedto the granules. After mixing, the mixture is compressed on a tabletmachine to afford a tablet weighing 100 mg.

Formulation Example E

Tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 25 mg Microcrystallinecellulose 400 mg Silicon dioxide fumed 10 mg Stearic acid 5 mg

Representative Procedure: The ingredients are thoroughly blended andthen compressed to form tablets (440 mg of composition per tablet).

Formulation Example F

Single-scored tablets for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 15 mg Cornstarch 50 mgCroscarmellose sodium 25 mg Lactose 120 mg Magnesium stearate 5 mg

Representative Procedure: The ingredients are thoroughly blended andcompressed to form a single-scored tablet (215 mg of compositions pertablet).

Formulation Example G

A suspension for oral administration is prepared as follows:

Ingredients Amount Compound of the invention 0.1 g Fumaric acid 0.5 gSodium chloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 gGranulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum k(Vanderbilt Co.) 1.0 g Flavoring 0.035 mL Colorings 0.5 mg Distilledwater q.s. to 100 mL

Representative Procedure: The ingredients are mixed to form a suspensioncontaining 10 mg of active ingredient per 10 mL of suspension.

Formulation Example H

A dry powder for administration by inhalation is prepared as follows:

Ingredients Amount Compound of the invention 1.0 mg Lactose 25 mg

Representative Procedure: The active ingredient is micronized and thenblended with lactose. This blended mixture is then loaded into a gelatininhalation cartridge. The contents of the cartridge are administeredusing a powder inhaler.

Formulation Example I

A dry powder for administration by inhalation in a metered dose inhaleris prepared as follows:

Representative Procedure: A suspension containing 5 wt. % of a compoundof the invention and 0.1 wt. % lecithin is prepared by dispersing 10 gof active compound as micronized particles with mean size less than 10μm in a solution formed from 0.2 g of lecithin dissolved in 200 mL ofdemineralized water. The suspension is spray dried and the resultingmaterial is micronized to particles having a mean diameter less than 1.5μm. The particles are loaded into cartridges with pressurized1,1,1,2-tetrafluoroethane.

Formulation Example J

An injectable formulation is prepared as follows:

Ingredients Amount Compound of the invention 0.2 g Sodium acetate buffersolution (0.4 M) 40 mL HCl (0.5 N) or NaOH (0.5 N) q.s. to pH 4 Water(distilled, sterile) q.s. to 20 mL

Representative Procedure: The above ingredients are blended and the pHis adjusted to 4±0.5 using 0.5 N HCl or 0.5 N NaOH.

Formulation Example K

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the Invention 4.05 mg Microcrystallinecellulose (Avicel PH 103) 259.2 mg Magnesium stearate 0.75 mg

Representative Procedure: The ingredients are thoroughly blended andthen loaded into a gelatin capsule (Size #1, White, Opaque) (264 mg ofcomposition per capsule).

Formulation Example L

Capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the Invention 8.2 mg Microcrystallinecellulose (Avicel PH 103) 139.05 mg Magnesium stearate 0.75 mg

Representative Procedure: The ingredients are thoroughly blended andthen loaded into a gelatin capsule (Size #1, White, Opaque) (148 mg ofcomposition per capsule).

It will be understood that any form of the compounds of the invention,(i.e. free base, pharmaceutical salt, or solvate) that is suitable forthe particular mode of administration, can be used in the pharmaceuticalcompositions discussed above.

Utility

The quinolinone-carboxamide compounds of the invention are 5-HT₄receptor agonists and therefore are expected to be useful for treatingmedical conditions mediated by 5-HT₄ receptors or associated with 5-HT₄receptor activity, i.e. medical conditions which are ameliorated bytreatment with a 5-HT₄ receptor agonist. Such medical conditionsinclude, but are not limited to, irritable bowel syndrome (IBS), chronicconstipation, functional dyspepsia, delayed gastric emptying,gastroesophageal reflux disease (GERD), gastroparesis, diabetic andidiopathic gastropathy, post-operative ileus, intestinalpseudo-obstruction, and drug-induced delayed transit. In addition, ithas been suggested that some 5-HT₄ receptor agonist compounds may beused in the treatment of central nervous system disorders includingcognitive disorders, behavioral disorders, mood disorders, and disordersof control of autonomic function.

In particular, the compounds of the invention increase motility of thegastrointestinal (GI) tract are thus are expected to be useful fortreating disorders of the GI tract caused by reduced motility inmammals, including humans. Such GI motility disorders include, by way ofillustration, chronic constipation, constipation-predominant irritablebowel syndrome (C-IBS), diabetic and idiopathic gastroparesis, andfunctional dyspepsia.

In one aspect, therefore, the invention provides a method of increasingmotility of the gastrointestinal tract in a mammal, the methodcomprising administering to the mammal a therapeutically effectiveamount of a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of the invention.

In another aspect, the invention provides a method of treating adisorder of reduced motility of the gastrointestinal tract in a mammal,the method comprising administering to the mammal, a therapeuticallyeffective amount of a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of the invention.

When used to treat disorders of reduced motility of the GI tract orother conditions mediated by 5-HT₄ receptors, the compounds of theinvention will typically be administered orally in a single daily doseor in multiple doses per day, although other forms of administration maybe used. The amount of active agent administered per dose or the totalamount administered per day will typically be determined by a physician,in the light of the relevant circumstances, including the condition tobe treated, the chosen route of administration, the actual compoundadministered and its relative activity, the age, weight, and response ofthe individual patient, the severity of the patient's symptoms, and thelike.

Suitable doses for treating disorders of reduced motility of the GItract or other disorders mediated by 5-HT₄ receptors will range fromabout 0.0007 to about 20 mg/kg/day of active agent, including from about0.0007 to about 1 mg/kg/day. For an average 70 kg human, this wouldamount to from about 0.05 to about 70 mg per day of active agent.

In one aspect of the invention, the compounds of the invention are usedto treat chronic constipation. When used to treat chronic constipation,the compounds of the invention will typically be administered orally ina single daily dose or in multiple doses per day. Preferably, the dosefor treating chronic constipation is expected to range from about 0.05to about 70 mg per day.

In another aspect of the invention, the compounds of the invention areused to treat irritable bowel syndrome. When used to treatconstipation-predominant irritable bowel syndrome, the compounds of theinvention will typically be administered orally in a single daily doseor in multiple doses per day. Preferably, the dose for treatingconstipation-predominant irritable bowel syndrome is expected to rangefrom about 0.05 to about 70 mg per day.

In another aspect of the invention, the compounds of the invention areused to treat diabetic gastroparesis. When used to treat diabeticgastroparesis, the compounds of the invention will typically beadministered orally in a single daily dose or in multiple doses per day.Preferably, the dose for treating diabetic gastroparesis is expected torange from about 0.05 to about 70 mg per day.

In yet another aspect of the invention, the compounds of the inventionare used to treat functional dyspepsia. When used to treat functionaldyspepsia, the compounds of the invention will typically be administeredorally in a single daily dose or in multiple doses per day. Preferably,the dose for treating functional dyspepsia is expected to range fromabout 0.05 to about 70 mg per day.

The invention also provides a method of treating a mammal having adisease or condition associated with 5-HT₄ receptor activity, the methodcomprising administering to the mammal a therapeutically effectiveamount of a compound of the invention or of a pharmaceutical compositioncomprising a compound of the invention.

Since the compounds of the invention are 5-HT₄ receptor agonists, suchcompounds also useful as research tools for investigating or studyingbiological systems or samples having 5-HT₄ receptors, or for discoveringnew 5-HT₄ receptor agonists. Moreover, since compounds of the inventionexhibit binding selectivity for 5-HT₄ receptors as compared with bindingto receptors of other 5-HT subtypes, particularly 5-HT₃ receptors, suchcompounds are particularly useful for studying the effects of selectiveagonism of 5-HT₄ receptors in a biological system or sample. Anysuitable biological system or sample having 5-HT₄ receptors may beemployed in such studies which may be conducted either in vitro or invivo. Representative biological systems or samples suitable for suchstudies include, but are not limited to, cells, cellular extracts,plasma membranes, tissue samples, mammals (such as mice, rats, guineapigs, rabbits, dogs, pigs, etc.) and the like.

In this aspect of the invention, a biological system or samplecomprising a 5-HT₄ receptor is contacted with a 5-HT₄ receptor-agonizingamount of a compound of the invention. The effects of agonizing the5-HT₄ receptor are then determined using conventional procedures andequipment, such as radioligand binding assays and functional assays.Such functional assays include ligand-mediated changes in intracellularcyclic adenosine monophosphate (cAMP), ligand-mediated changes inactivity of the enzyme adenylyl cyclase (which synthesizes cAMP),ligand-mediated changes in incorporation of analogs of guanosinetriphosphate (GTP), such as [³⁵S]GTPγS (guanosine5′-O-(γ-thio)triphosphate) or GTP-Eu, into isolated membranes viareceptor catalyzed exchange of GTP analogs for GDP analogs,ligand-mediated changes in free intracellular calcium ions (measured,for example, with a fluorescence-linked imaging plate reader or FLIPR®from Molecular Devices, Inc.), and measurement of mitogen activatedprotein kinase (MAPK) activation. A compound of the invention mayagonize or increase the activation of 5-HT₄ receptors in any of thefunctional assays listed above, or assays of a similar nature. A 5-HT₄receptor-agonizing amount of a compound of the invention will typicallyrange from about 1 nanomolar to about 1000 nanomolar.

Additionally, the compounds of the invention can be used as researchtools for discovering new 5-HT₄ receptor agonists. In this embodiment,5-HT₄ receptor binding or functional data for a test compound or a groupof test compounds is compared to the 5-HT₄ receptor binding orfunctional data for a compound of the invention to identify testcompounds that have superior binding or functional activity, if any.This aspect of the invention includes, as separate embodiments, both thegeneration of comparison data (using the appropriate assays) and theanalysis of the test data to identify test compounds of interest.

Among other properties, compounds of the invention have been found to bepotent agonists of the 5-HT₄ receptor and to exhibit substantialselectivity for the 5-HT₄ receptor subtype over the 5-HT₃ receptorsubtype in radioligand binding assays. Further, compounds of theinvention which have been tested in a rat model have typicallydemonstrated superior pharmacokinetic properties in a rat model.Compounds of the invention are thus expected to be bioavailable uponoral administration. In addition, these compounds typically have beenshown to exhibit an acceptable level of inhibition of the potassium ioncurrent in an in vitro voltage-clamp model using isolated whole cellsexpressing the hERG cardiac potassium channel. The voltage-clamp assayis an accepted pre-clinical method of assessing the potential forpharmaceutical agents to change the pattern of cardiac repolarization,specifically to cause, so-called QT prolongation, which has beenassociated with cardiac arrhythmia. (Cavero et al., Opinion onPharmacotherapy, 2000, 1, 947-73, Fermini et al., Nature Reviews DrugDiscovery, 2003, 2, 439-447) Accordingly, pharmaceutical compositionscomprising compounds of the invention are expected to have an acceptablecardiac profile.

These properties, as well as the utility of the compounds of theinvention, can be demonstrated using various in vitro and in vivo assayswell-known to those skilled in the art. Representative assays aredescribed in further detail in the following examples.

Examples

The following synthetic and biological examples are offered toillustrate the invention, and are not to be construed in any way aslimiting the scope of the invention. In the examples below, thefollowing abbreviations have the following meanings unless otherwiseindicated. Abbreviations not defined below have their generally acceptedmeanings.

-   -   Boc=tert-butoxycarbonyl    -   (Boc)₂O=di-tert-butyl dicarbonate    -   DCM=dichloromethane    -   DMF=N,N-dimethylformamide    -   DMSO=dimethyl sulfoxide    -   EtOAc=ethyl acetate    -   mCPBA=m-chloroperbenzoic acid    -   MeCN=acetonitrile    -   MTBE=tert-butyl methyl ether    -   PyBOP=benzotriazol-1-yloxytripyrrolidino-phosphonium        hexafluorophosphate    -   R_(f)=retention factor    -   RT=room temperature    -   TFA=trifluoroacetic acid    -   THF=tetrahydrofuran

Reagents and solvents were purchased from commercial suppliers (Aldrich,Fluka, Sigma, etc.), and used without further purification. Reactionswere run under nitrogen atmosphere, unless noted otherwise. Progress ofreaction mixtures was monitored by thin layer chromatography (TLC),analytical high performance liquid chromatography (anal. HPLC), and massspectrometry, the details of which are given below and separately inspecific examples of reactions. Reaction mixtures were worked up asdescribed specifically in each reaction; commonly they were purified byextraction and other purification methods such as temperature-, andsolvent-dependent crystallization, and precipitation. In addition,reaction mixtures were routinely purified by preparative HPLC: a generalprotocol is described below. Characterization of reaction products wasroutinely carried out by mass and ¹H-NMR spectrometry. For NMRmeasurement, samples were dissolved in deuterated solvent (CD₃OD, CDCl₃,or DMSO-d₆), and ¹H-NMR spectra were acquired with a Varian Gemini 2000instrument (300 MHz) under standard observation conditions. Massspectrometric identification of compounds was performed by anelectrospray ionization method (ESMS) with a Perkin Elmer instrument (PESCIEX API 150 EX).

General Protocol for Analytical HPLC

Crude compounds were dissolved in 50% MeCN/H₂O (with 0.1% TFA) at0.5-1.0 mg/mL concentration, and analyzed using the followingconditions:

-   -   Column: Zorbax Bonus-RP (3.5 μm of particle size, 2.1×50 mm)    -   Flow rate: 0.5 mL/min    -   Mobile Phases: 5% MeCN/H₂O containing 0.1% TFA (isocratic; 0-0.5        min);        -   5% MeCN/H₂O containing 0.1% TFA to 75% MeCN/H₂O containing            0.1% TFA (linear gradient 0.5-4 min);    -   Detector wavelength: 214, 254, and 280 nm.        Other conditions, when used, are indicated explicitly.        General Protocol for Preparative HPLC Purification

Crude compounds were dissolved in 50% acetic acid in water at 50-100mg/mL concentration, filtered, and fractionated using the followingprocedure:

-   -   Column: YMC Pack-Pro C18 (50a×20 mm; ID=5 μm)    -   Flow rate: 40 mL/min    -   Mobile Phases: A=90% MeCN/10% H₂O/0.1% TFA        -   B=98% H₂O/2% MeCN/0.1% TFA    -   Gradient: 10% A/90% B to 50% A/50% B over 30 min (linear)    -   Detector wavelength: 214 nm.        Preparation of Secondary Amines

Preparation of various secondary amines used as intermediates in thesynthesis of a compound of formula (I) are described below.

Thiomorpholine-1,1-dioxide was prepared from thiomorpholine byprotection of the secondary amine to N-Boc thiomorpholine ((Boc)₂O,MeOH), oxidation to sulfone (mCPBA, CH₂Cl₂, 0° C.), and deprotection ofthe N-Boc group to provide the free amine (CF₃CO₂H, CH₂Cl₂). (m/z):[M+H]⁺ calcd for C₄H₉NO₂S, 136.04; found, 135.9.

The N-sulfonyl derivatives of piperazine were prepared from N-Bocpiperazine by reacting with respective sulfonyl chloride (iPr₂NEt,CH₂Cl₂, 0° C.), and deprotecting the N-Boc group (CF₃CO₂H, CH₂Cl₂).1-Methanesulfonylpiperazine: ¹H-NMR (CDCl₃; neutral): δ (ppm) 3.1 (t,4H), 2.9 (t, 4H), 2.7 (s, 3H).1-(Methylsulfonyl)methanesulfonyl-piperazine: ¹H-NMR (CD₃OD): δ (ppm)2.90 (s, 3H), 3.02 (m, 4H), 3.38 (m, 4H), 4.61 (s, 2H).Methanesulfonylpiperazine was also prepared by reacting methanesulfonylchloride with excess piperazine (>2 equivalents) in water.

The racemic or single chiral isomer forms of 3-acetylaminopyrrolidinewere prepared by treating N¹-Boc-3-aminopyrrolidine (racemate, 3R, or3S) with acetyl chloride (iPr₂NEt, CH₂Cl₂, 0° C.), and deprotecting theN-Boc group (CF₃CO₂H, CH₂Cl₂). 3-(Acetamido)pyrrolidine: ¹H-NMR(DMSO-d₆; TFA salt): δ (ppm) 4.2 (quin, 1H), 3.3-3.1 (m, 3H), 2.9 (m,1H), 2.0 (m, 1H), 1.8 (br s, 4H).

3-((R)-2-Hydroxypropionamido)pyrrolidine was prepared after amidation ofN¹-Boc-3-aminopyrrolidine (L-lactic acid, PyBOP, DMF, RT), anddeprotection of N-Boc group (CF₃CO₂H, CH₂Cl₂). (m/z): [M+H]⁺ calcd forC₇H₁₄N₂O₂, 159.11; found, 159.0. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 4.4(quin, 1H), 4.1 (q, 1H), 3.5-3.4 (m, 2H), 3.3-3.2 (m, 2H), 2.3 (m, 1H),2.0 (m, 1H), 1.3 (d, 3H).

The N³-alkanesulfonyl derivatives of (3R)-aminopyrrolidine were obtainedby treating N¹-Boc-(3R)-aminopyrrolidine with propionylsulfonyl chlorideor cyclohexylmethylsulfonyl chloride (i-Pr₂NEt, CH₂Cl₂, 0° C.), anddeprotecting N-Boc group (CF₃CO₂H, CH₂Cl₂).

3-(N-Acetyl-N-methylamido)piperidine was prepared from N³-Cbz protected3-amino-piperidine-1-carboxylic acid t-butyl ester (De Costa, B., et al.J. Med. Chem. 1992, 35, 4334-43) after four synthetic steps: i) MeI,n-BuLi, THF, -78° C. to rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) AcCl,i-Pr₂NEt, CH₂Cl₂; iv) CF₃CO₂H, CH₂Cl₂. m/z: [M+H]⁺ calcd for C₈H₁₆N₂O:157.13; found, 157.2. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 4.6 (m, 1H), 3.3(m, 1H), 3.2 (m, 1H), 3.0 (m, 1H), 2.9 (s, 3H), 2.8 (m, 1H), 2.0 (s,3H), 1.9-1.7 (m, 4H).

3-(N-Acetyl-amido)piperidine was prepared from3-amino-piperidine-1-carboxylic acid tert-butyl ester afterN-acetylation and deprotection of the N-Boc group: i) AcCl, i-Pr₂NEt,CH₂Cl₂; ii) CF₃CO₂H, CH₂Cl₂. ¹H-NMR (CD₃OD; TFA salt): δ (ppm) 3.9 (m,1H), 3.3 (dd, 1H), 3.2 (m, 1H), 2.9 (dt, 1H), 2.75 (dt, 1H), 2.0-1.9 (m,2H), 1.9 (s, 3H), 1.8-1.4 (m, 2H).

The N³-alkanesulfonyl derivatives of 3-aminopiperidine were synthesizedby reacting the chiral or racemic forms of3-amino-piperidine-1-carboxylic acid tert-butyl ester with therespective alkanesulfonyl chloride (i-Pr₂NEt, CH₂Cl₂) and deprotectingthe N-Boc group (CF₃CO₂H, CH₂Cl₂).(3S)-3-(ethanesulfonylamido)piperidine: ¹H-NMR (CD₃OD): δ (ppm) 1.29 (t,3H, J₁=7.4 Hz), 1.50-1.80 (m, 2H), 1.90-2.10 (m, 2H), 2.89 (m, 2H), 3.05(q, 2H, J₁=7.4 Hz), 3.27 (m, 2H), 3.40 (d of d(br), 1H), 3.52 (m, 1H).3S-Methylsulfonylmethanesulfonylamido-piperidine: ¹H-NMR (CD₃OD): δ(ppm) 2.13-2.30 (m, 2H), 2.40-2.57 (m, 2H), 2.98 (m, 2H), 3.15 (s, 3H),3.21 (m, 2H), 3.30 (br d, 1H), 3.74 (m, 1H).

3-(Methylamino)-1-acetylpyrrolidine was prepared from3-(methylamino)-1-benzylpyrrolidine (TCI America) after four steps: i)(Boc)₂O, MeOH, rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) AcCl, i-Pr₂NEt,CH₂Cl₂; iv) CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd for C₇H₁₄N₂O: 143.12;found, 143.0.

3-(Methylamino)-1-(methanesulfonyl)pyrrolidine was prepared from3-(methylamino)-1-benzylpyrrolidine after four steps: i) (Boc)₂O, MeOH,rt; ii) H₂ (1 atm), 10% Pd/C, EtOH; iii) CH₃SO₂Cl, i-Pr₂NEt, CH₂Cl₂; iv)CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd for C₆H₁₄N₂O₂S: 179.08; found,179.2. 3R-Methylamino-1-(methanesulfonyl)pyrrolidine was prepared in asimilar manner from (3R)-(methylamino)-1-benzylpyrrolidine.

Derivatives of tetrahydro-3-thiophenamine-1,1-dioxide were preparedfollowing the protocol of Loev, B. J. Org. Chem. 1961, 26, 4394-9 byreacting 3-sulfolene with a requisite primary amine in methanol (cat.KOH, rt). N-Methyl-3-tetrahydrothiopheneamine-1,1-dioxide (TFA salt):¹H-NMR (DMSO-d₆): δ (ppm) 9.4 (br s, 2H), 4.0-3.8 (quin, 1H), 3.6-3.5(dd, 1H), 3.4-3.3 (m, 1H), 3.2-3.1 (m, 2H), 2.5 (s, 3H), 2.4 (m, 1H),2.1 (m, 1H). N²-(1-hydroxy)ethyl-3-tetrahydrothiopheneamine-1,1-dioxide:(m/z): [M+H]⁺ calcd for C₆H₁₃NO₃S: 180.07; found, 180.2.

(S)-1,1-Dioxo-tetrahydro-1λ⁶-thiophen-3-ylamine was prepared asfollows: 1) N-Boc protection of (S)-3-tetrahydrothiophenamine (Dehmlow,E. V.; Westerheide, R. Synthesis 1992, 10, 947-9) by treating with(Boc)₂O in methanol at room temperature for about 12 h; 2) oxidation bytreating with mCPBA in dichloromethane to N-Boc protected(S)-1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-ylamine at 0° C. for about 5 h;and 3) N-Boc deprotection of the sulfone derivative with TFA indichloromethane at room temperature for 1 h to the free amine which wasisolated as a TFA salt. (R)-1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-ylaminewas prepared using the same method, but replacing the(S)-3-tetra-hydrothiophenamine with (R)-3-tetrahydrothiophenamine.

N-Methyl-tetrahydro-2H-thiopyran-4-amine-1,1-dioxide was prepared fromtetrahydro-4H-thiopyran-4-one: i) MeNH₂, NaBH₄; ii) (Boc)₂O, MeOH; iii)mCPBA, CH₂Cl₂, 0° C.; iv) CF₃CO₂H, CH₂Cl₂. (m/z): [M+H]⁺ calcd forC₆H₁₃NO₂S 164.07; found, 164.9. ¹H-NMR (CD₃OD; TFA salt): δ (ppm)3.4-3.1 (m, 5H), 2.7 (s, 3H), 2.4 (br d, 2H), 2.1 (br m, 2H).

1-Acetyl-3-(methylamino)piperidine was prepared from N³-Cbz protected3-methylamino-piperidine: i) AcCl, i-Pr₂NEt, CH₂Cl₂; ii) H₂ (1 atm), 10%Pd/C, EtOH. ¹H-NMR (CD₃OD): δ (ppm) 4.0 (m, 1H), 3.6 (m, 1H), 3.4-3.2(m, 2H), 3.0 (m, 1H), 2.6 (s, 3H), 2.1 (s, 3H), 1.8-1.6 (m, 4H).

1-(Methanesulfonyl)-3-(methylamino)piperidine was prepared from N³-Cbzprotected 3-methylamino-piperidine: i) CH₃SO₂Cl, i-Pr₂NEt, CH₂Cl₂; ii)H₂ (1 atm), 10% Pd/C, EtOH. (m/z): [M+H]⁺ calcd for C₇H₁₆N₂O₂S 193.10;found, 193.0. ¹H-NMR (DMSO-d₆; TFA salt): δ (ppm) 3.4 (dd, 1H), 3.2 (m,2H), 3.10 (s, 3H), 3.0-2.9 (m, 2H), 2.8 (s, 3H), 1.85-1.75 (m, 2H),1.6-1.4 (m, 2H).

Proline dimethylamide, and iminodiacetonitrile were purchased fromBachem, and Aldrich, respectively.

The N-derivatives of piperazine such as 1-(methoxycarbonyl)piperazine,1-(dimethylaminocarbonyl)piperazine, and1-(dimethylaminosulfonyl)piperazine were prepared by reacting piperazinewith methylchloroformate, dimethylaminochoroformate, ordimethylaminosulfamoyl chloride, respectively.

1-Methylamino-2-methylsulfonylethane was obtained by reactingmethylamine with methyl vinyl sulfone in methanol.N-[2-(2-methoxyethylamino)ethyl], N-methyl-methanesulfonamide wassynthesized starting from partially N-Boc protected ethanediamine afterfour steps of reactions in a sequence as follows: i) methylsulfonylchloride, triethylamine; ii) MeI, Cs₂CO₃; iii) NaH,1-bromo-2-methoxyethane; iv) CF₃CO₂H.

Isonipecotamide (piperidine-4-carboxamide), and proline amide werepurchased from Aldrich. 2-Hydroxymethylmorpholine was available fromTyger Scientific Product.

Methyl 4-piperidinylcarbamate was prepared from the reaction of N₁-Bocprotected 4-aminopiperidine with methylchloroformate followed by thedeprotection of the N-Boc group.

4-Piperidinol-dimethylcarbamate, and N-dimethyl-N-(3-piperidinyl)ureawere prepared by reacting dimethylcarbamoyl chloride with N-Bocprotected 4-piperidinol or N₁-Boc-3-aminopiperidine, respectively.

3-(Methylamino)-1-(dimethylaminosulfonyl)pyrrolidine was obtained byreacting 3-(N-methyl-N-Boc-amino)pyrrolidine with dimethylsulfamoylchloride.

2-(3-Pyrrolidinyl)isothiazolidine-1,1-dioxide was synthesized bytreating N₁-Boc protected 3-aminopyrrolidine with 3-chloropropylsulfonylchloride in the presence of triethylamine, and followed by TFA treatmentfor the deprotection of the Boc group.

Example 1 Synthesis of(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)carbamicacid methyl ester

a. Preparation of 8-benzyl-8-azabicyclo[3.2.1]octan-3-one

Concentrated hydrochloric acid (30 mL) was added to a heterogeneoussolution of 2,5-dimethoxy tetrahydrofuran (82.2 g, 0.622 mol) in water(170 mL) while stirring. In a separate flask cooled to 0° C. (ice bath),concentrated hydrochloric acid (92 mL) was added slowly to a solution ofbenzyl amine (100 g, 0.933 mol) in water (350 mL). The2,5-dimethoxytetrahydrofuran solution was stirred for approximately 20min, diluted with water (250 mL), and then the benzyl amine solution wasadded, followed by the addition of a solution of 1,3-acetonedicarboxylicacid (100 g, 0.684 mol) in water (400 mL) and then the addition ofsodium hydrogen phosphate (44 g, 0.31 mol) in water (200 mL). The pH wasadjusted from pH 1 to pH ˜4.5 using 40% NaOH. The resulting cloudy andpale yellow solution was stirred overnight. The solution was thenacidified to pH 3 from pH 7.5 using 50% hydrochloric acid, heated to 85°C. and stirred for 2 hours. The solution was cooled to room temperature,basified to pH 12 using 40% NaOH, and extracted with dichloromethane(3×500 mL). The combined organic layers were washed with brine, dried(MgSO₄), filtered and concentrated under reduced pressure to produce thecrude title intermediate as a viscous brown oil (52 g).

To a solution of the crude intermediate in methanol (1000 mL) was addeddi-tert-butyl dicarbonate (74.6 g, 0.342 mol) at 0° C. The solution wasallowed to warm to room temperature and stirred overnight. The methanolwas removed under reduced pressure and the resulting oil was dissolvedin dichloromethane (1000 mL). The intermediate was extracted into 1 MH₃PO₄ (1000 mL) and washed with dichloromethane (3×250 mL). The aqueouslayer was basified to pH 12 using aqueous NaOH, and extracted withdichloromethane (3×500 mL). The combined organic layers were dried(MgSO₄), filtered and concentrated under reduced pressure to produce thetitle intermediate as a viscous, light brown oil. ¹H-NMR (CDCl₃) δ (ppm)7.5-7.2 (m, 5H, C₆H₅), 3.7 (s, 2H, CH₂Ph), 3.45 (broad s, 2H, CH—NBn),2.7-2.6 (dd, 2H, CH₂CO), 2.2-2.1 (dd, 2H, CH₂CO), 2.1-2.0 (m, 2H,CH₂CH₂), 1.6 (m, 2H, CH₂CH₂). (m/z): [M+H]⁺ calcd for C₁₄H₁₇NO 216.14;found, 216.0.

b. Preparation of 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester

To a solution of 8-benzyl-8-azabicyclo[3.2.1]octan-3-one (75 g, 0.348mol) in EtOAc (300 mL) was added a solution of di-tert-butyl dicarbonate(83.6 g, 0.383 mol, 1.1 eq) in EtOAc (300 mL). The resulting solutionand rinse (100 mL EtOAc) was added to a 1 L Parr hydrogenation vesselcontaining 23 g of palladium hydroxide (20 wt. % Pd, dry basis, oncarbon, ˜50% wet with water; e.g. Pearlman's catalyst) under a stream ofnitrogen. The reaction vessel was degassed (alternating vacuum and N₂five times) and pressurized to 60 psi of H₂ gas. The reaction solutionwas agitated for two days and recharged with H₂ as needed to keep the H₂pressure at 60 psi until the reaction was complete as monitored bysilica thin layer chromatography. The solution was then filtered througha pad of Celite® and concentrated under reduced pressure to yield thetitle intermediate quantitatively as a viscous, yellow to orange oil (51g). It was used in the next step without further treatment. ¹H NMR(CDCl₃)

(ppm) 4.5 (broad, 2H, CH—NBoc), 2.7 (broad, 2H, CH₂CO), 2.4-2.3 (dd, 2H,CH₂CH₂), 2.1 (broad m, 2H, CH₂CO), 1.7-1.6 (dd, 2H, CH₂CH₂), 1.5 (s, 9H,(CH₃)₃COCON)).

c. Preparation of(1S,3R,5R)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester

To a solution of the product of the previous step (75.4 g, 0.335 mol) inmethanol (1 L) was added ammonium formate (422.5 g, 6.7 mol), water (115mL) and 65 g of palladium on activated carbon (10% on dry basis, 50% wetwith water; Degussa type E101NE/W) under a stream of N₂ while stirringvia mechanical stirrer. After 24 and 48 hours, additional portions ofammonium formate (132 g, 2.1 mol) were added each time. Once reactionprogression ceased, as monitored by anal. HPLC, Celite® (>500 g) wasadded and the resulting thick suspension was filtered and then thecollected solid was rinsed with methanol (˜500 mL). The filtrates werecombined and concentrated under reduced pressure until all methanol hadbeen removed. The resulting cloudy, biphasic solution was then dilutedwith 1M phosphoric acid to a final volume of 1.5 to 2.0 L at pH 2 andwashed with dichloromethane (3×700 mL). The aqueous layer was basifiedto pH 12 using 40% aq. NaOH, and extracted with dichloromethane (3×700mL). The combined organic layers were dried over MgSO₄, filtered, andconcentrated by rotary evaporation, then high-vacuum leaving 52 g (70%)of the title intermediate, commonly N-Boc-endo-3-aminotropane, as awhite to pale yellow solid. The isomer ratio of endo to exo amine of theproduct was >99:1 based on ¹H-NMR analysis (>96% purity by analyticalHPLC). ¹H NMR (CDCl₃) δ (ppm) 4.2-4.0 (broad d, 2H, CHNBoc), 3.25 (t,1H, CHNH₂), 2.1-2.05 (m, 4H), 1.9 (m, 2H), 1.4 (s, 9H, (CH₃)₃OCON),1.2-1.1 (broad, 2H). (m/z): [M+H]⁺ calcd for C₁₂H₂₂N₂O₂) 227.18; found,227.2. Analytical HPLC (isocratic method; 2:98 (A:B) to 90:10 (A:B) over5 min): retention time=3.68 min.

d. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid

Acetone (228.2 mL, 3.11 mol) was added to a stirred suspension of2-aminophenylmethanol (255.2 g, 2.07 mol) and acetic acid (3.56 mL, 62mmol) in water (2 L) at room temperature. After 4 h, the suspension wascooled to 0° C. and stirred for an additional 2.5 h and then filtered.The solid was collected and washed with water and the wet solid cooledand dried by lyophilisation to yield2,2,-dimethyl-1,4-dihydro-2H-benzo[1,3]oxazine (332.2 g, 98%) as anoff-white solid. ¹H NMR (CDCl₃; 300 MHz): 1.48 (s, 6H, C(CH ₃)₂), 4.00(bs, 1H, NH), 4.86 (s, 2H, CH ₂), 6.66 (d, 1H, ArH), 6.81 (t, 1H, ArH),6.96 (d, 1H, ArH), 7.10 (t, 1H, ArH).

A solution of 2,2,-dimethyl-1,4-dihydro-2H-benzo[1,3]oxazine (125 g,0.77 mol) in THF (1 L) was filtered through a scintillation funnel andthen added dropwise via an addition funnel, over a period of 2.5 h, to astirred solution of 1.0 M LiAlH₄ in THF (800 mL) at 0° C. The reactionwas quenched by slow portionwise addition of Na₂SO₄.10H₂O (110 g), overa period of 1.5 h, at 0° C. The reaction mixture was stirred overnight,filtered and the solid salts were washed thoroughly with THF. Thefiltrate was concentrated under reduced pressure to yield2-isopropylaminophenylmethanol (120 g, 95%) as a yellow oil. ¹H NMR(CDCl₃; 300 MHz): 1.24 (d, 6H, CH(CH ₃)₂), 3.15 (bs, 1H, OH), 3.61(sept, 1H, CH(CH₃)₂), 4.57 (s, 2H, CH ₂), 6.59 (t, 1H, ArH), 6.65 (d,1H, ArH), 6.99 (d, 1H, ArH), 7.15 (t, 1H, ArH).

Manganese dioxide (85% 182.6 g, 1.79 mol) was added to a stirredsolution of 2-isopropylaminophenylmethanol (118 g, 0.71 mol) in toluene(800 mL) and the reaction mixture was heated to 117° C. for 4 h. Thereaction mixture was allowed to cool to room temperature overnight andthen filtered through a pad of Celite which was eluted with toluene. Thefiltrate was concentrated under reduced pressure to yield2-isopropylaminobenzaldehyde (105 g, 90%) as an orange oil. ¹H NMR(CDCl₃; 300 MHz): 1.28 (d, 6H, CH(CH ₃)₂), 3.76 (sept, 1H, CH(CH₃)₂),6.65 (t, 1H, ArH), 6.69 (d, 1H, ArH), 7.37 (d, 1H, ArH), 7.44 (t, 1H,ArH), 9.79 (s, 1H, CHO).

2,2-Dimethyl-[1,3]dioxane-4,6-dione, commonly Meldrum's acid, (166.9 g,1.16 mol) was added to a stirred solution of2-isopropylaminobenzaldehyde (105 g, 0.64 mol), acetic acid (73.6 mL,1.29 mol) and ethylenediamine (43.0 mL, 0.64 mol) in methanol (1 L) at0° C. The reaction mixture was stirred for 1 h at 0° C. and then at roomtemperature overnight. The resulting suspension was filtered and thesolid washed with methanol and collected to yield the titleintermediate, 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid(146 g, 98%) as an off-white solid. ¹H NMR (CDCl₃; 300 MHz): 1.72 (d,6H, CH(CH ₃)₂), 5.50 (bs, 1H, CH(CH₃)₂), 7.44 (t, 1H, ArH), 7.75-7.77(m, 2H, ArH), 7.82 (d, 1H, ArH), 8.89 (s, 1H, CH).

e. Preparation of(1S,3R,5R)-3-[1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester

Thionyl chloride (36.6 mL, 0.52 mol) was added to a stirred suspensionof 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (80 g, 0.35mol) in toluene (600 mL) at 85° C. and the reaction mixture then heatedto 95° C. for 2 h. The reaction mixture was cooled to room temperatureand then added over 25 min to a vigorously stirred biphasic solution of(1S,3R,5R)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester (78.2 g, 0.35 mol) and sodium hydroxide (69.2 g, 1.73mol) in toluene/water (1:1) (1 L) at ° C. After 1 h, the layers wereallowed to separate and the organic phase concentrated under reducedpressure. The aqueous phase was washed with EtOAc (1 L) and then (500mL) and the combined organic extracts used to dissolve the concentratedorganic residue. This solution was washed with 1M H₃PO₄ (500 mL),saturated aqueous NaHCO₃ (500 mL) and brine (500 mL), dried over MgSO₄,filtered and concentrated under reduced pressure to yield the titleintermediate (127.9 g, approx. 84%) as a yellow solid. ¹H NMR (CDCl₃):1.47 (s, 9H), 1.67 (d, 6H), 1.78-1.84 (m, 2H), 2.04-2.18 (m, 6H),4.20-4.39 (m, 3H), 5.65 (bs, 1H), 7.26 (dd. 1H), 7.63 (m, 2H), 7.75 (dd,1H), 8.83 (s, 1H), 10.63 (d, 1H).

f. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid {(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide

TFA (300 mL) was added to a stirred solution of the product of theprevious step (127.9 g) in CH₂Cl₂ (600 mL) at 0° C. The reaction mixturewas warmed to room temperature and stirred for 1 h and then concentratedunder reduced pressure. The oily brown residue was then poured into avigorously stirred solution of ether (3 L) and a solid precipitateformed immediately. The suspension was stirred overnight and then thesolid collected by filtration and washed with ether to yield the titleintermediate as its trifluoroacetic acid salt (131.7 g, 86% over twosteps) as a light yellow solid. ¹H NMR (CDCl₃): 1.68 (d, 6H), 2.10 (d,2H), 2.33-2.39 (m, 4H), 2.44-2.61 (m, 2H), 4.08 (bs, 2H), 4.41 (m, 1H),5.57 (bs, 1H), 7.31 (m. 1H), 7.66 (m, 2H), 7.77 (d, 1H), 8.83 (s, 1H),9.38 (bd, 2H), 10.78 (d, 1H).

g. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{(1S,3R,5R)-8-[(2,2-dimethoxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

N,N′-diisopropylethylamine (4.3 mL) and dimethoxyacetaldehyde intert-butyl methyl ether (conc 45%; 4.5 mL, 17 mmol) were added to asolution of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide mono trifluoroacetic acidsalt (5.44 g; 12 mmol) dissolved in 50 mL of dichloromethane. Afterstirring 35 minutes at ambient temperature, sodium triacetoxyborohydride(3.7 g; 17.3 mmol) was added to the reaction mixture. After 90 minutes,water (50 mL) and saturated NaHCO₃ solution (100 mL) was slowly added tothe reaction mixture in an ice bath to quench the reaction. The mixturewas diluted with 500 mL of dichloromethane, and transferred to aseparatory funnel. The organic layer was collected, and washed withsaturated NaHCO₃ (250 mL), and brine solution (350 mL). It was driedover MgSO₄, and evaporated in vacuo, to yield the title intermediate.(m/z): [M+H]⁺ calcd for C₂₄H₃₃N₃O₄ 428.25; found, 428.4.

h. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{(1S,3R,5R)-8-[(2,2-dihydroxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

1-Isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[(2,2-dimethoxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide(5.5 g) was suspended in 50 mL of 6M hydrochloric acid, then heated at70° C. for 1 h. The reaction mixture was cooled to 0° C., and dilutedwith dichloromethane (100 mL) prior to basification of the aqueous layerby slow addition of 6M NaOH (80 mL). It was further mixed with 80 mL ofdichloromethane, and transferred to a separatory funnel. The organiclayer was collected, washed with brine, dried over MgSO₄, and evaporatedto dryness to yield the title intermediate as an aldehyde hydrate.(m/z): [M+H]⁺ calcd for C₂₂H₂₉N₃O₄ 400.22; found, 400.5.

i. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{(1S,3R,5R)-{8-[2-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}-amide

1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamine trifluoroacetic acid salt(500 mg; 2 mmol), N,N′-diisopropylethylamine (0.35 mL), and sodiumtriacetoxyborohydride (422 mg; 2 mmol) was added to a vial containing 10mL of dichloromethane. The mixture was stirred for 5 minutes prior tothe addition of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[(2,2-dihydroxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide(0.427 g). The final mixture was stirred for 1 h, at which time thereaction was judged to be complete based on HPLC and mass spectrometricanalysis. Water (20 mL) was slowly added to quench the remainingreducing agent. The mixture was diluted with 100 mL of dichloromethane,and shaken in a funnel before collecting the organic layer. The organiclayer was washed with 1M NaOH (40 mL) and brine (50 mL), dried overMgSO₄, and evaporated to yield the title intermediate as a colorlesssolid. This crude product was used in the next step without furthertreatment.

j. Synthesis of(1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-aza-bicyclo[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester

N,N′-diisopropylethylamine (0.07 mL, 0.4 mmol) and methyl chloroformate(0.02 mL, 0.26 mmol) was added to a solution of DMF (1 mL) containingthe product of the previous step,1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-{8-[2-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}-amide(65 mg, 0.13 mmol). The reaction mixture was shaken at room temperaturefor 30 minutes, and concentrated in vacuo, yielding an oily residue. Theresidue was dissolved in 50% aqueous acetic acid (1 mL), and purified bypreparative HPLC, to yield the title compound. (m/z): [M+H]⁺ calcd forC₂₈H₃₈N₄O₆S 559.25; found, 559.2. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.56 min.

Example 2 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)methanesulfonylamino]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

Following the procedure described in Example 1, step (j), the titlecompound was prepared by replacing methyl chloroformate,N,N′-diisopropylethylamine, and DMF with methylsulfonyl chloride, DBU,and dichloromethane, respectively. (m/z): [M+H]⁺ calcd for C₂₇H₃₈N₄O₆S₂579.22; found. 579.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.62 min.

Example 3 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3,3-dimethylureido]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

Following the procedure described in Example 1, step (j), the titlecompound was prepared by replacing methyl chloroformate withN,N′-dimethylcarbamoyl chloride. (m/z): [M+H]⁺ calcd for C₂₉H₄₁N₅O₅S572.28; found, 572.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.55 min.

Example 4 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3-methylureido]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

Following the procedure described in Example 1, step (j), the titlecompound was prepared by replacing methyl chloroformate with methylisocyanate. (m/z): [M+H]⁺ calcd for C₂₈H₃₉N₅O₅S 558.27; found 558.2[M+H]⁺. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.69min.

Example 5 Synthesis of(1,1-dioxohexahydro-1λ⁶-thiopyran-4-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester

Following the procedure described in Example 1, the title compound wasprepared by replacing 1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylaminetrifluoroacetic acid salt in Example 1, step (i), with1,1-dioxohexahydro-1λ⁶-thiopyran-4-ylamine. (m/z): [M+H]⁺ calcd forC₂₉H₄₀N₄O₆S 573.27; found, 573.2. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.70 min.

Example 6 Synthesis of((R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-ethyl)carbamicacid methyl ester

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{8-(1S,3R,5R)-[2-((R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)ethyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide

(R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamine trifluoroacetic acid salt(278 mg; 1.1 mmol), and sodium triacetoxyborohydride (254 mg; 1.2 mmol)was added to a vial containing 4 mL of dichloromethane. The mixture wasstirred for 5 min prior to the addition of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[(2,2-dihydroxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide(0.420 g; 1.1 mmol), the product of Example 1, step (h). The mixture wasstirred for 1 h, at which time the reaction was judged to be completebased on HPLC and mass spectrometric analysis. Water (10 mL) was slowlyadded to quench the remaining reducing agent. The mixture was dilutedwith 50 mL of dichloromethane, and shaken in a funnel before collectingthe organic layer. It was washed with 1M NaOH (20 mL) and brine (20 mL),dried over MgSO₄, and evaporated to yield the title intermediate as acolorless solid. This crude product was used in the next step withoutfurther purification. (m/z): [M+H]⁺ calcd for C₂₆H₃₆N₄O₄S calcd. 501.25;found, 501.6.

b. Synthesis of((R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-ethyl)carbamicacid methyl ester

N,N′-diisopropylethylamine (0.38 mL, 2.2 mmol) and methyl chloroformate(0.11 mL, 1.4 mmol) was added to a solution of DMF (1 mL) containing1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{8-(1S,3R,5R)-[2-((R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)ethyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide(365 mg, 0.73 mmol) prepared in step (a) above. The reaction mixture wasshaken at room temperature for about 30 minutes, then concentrated invacuo, yielding an oily residue. The residue was dissolved in 50%aqueous acetic acid (1 mL), and purified by preparative HPLC, to yieldthe title compound. (m/z): [M+H]⁺ calcd for C₂₈H₃₈N₄O₆S 559.27; found,559.4. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.56 min.

Example 7 Synthesis of((S)-1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-ethyl)carbamicacid methyl ester

The title compound, an (S)-enantiomer of the compound of Example 6, wasprepared using the process described in Example 6, by replacing inExample 6, step (a), (R)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamine with(S)-1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamine. (m/z): [M+H]⁺ calcd forC₂₈H₃₈N₄O₆S 559.27; found, 559.4. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.56 min.

Example 8 Synthesis of(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-(3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamicacid methyl ester

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid [8-(1S,3R,5R)-(3-aminopropyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

N,N′-diisopropylethylamine (15.7 mL, 90 mmol), andN-Boc-3-bromo-propanamine (14.2 g, 60 mmol) was added to a solution of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide mono trifluoroacetic acidsalt (13.6 g; 30 mmol) (the product of Example 1, step (f)) dissolved in120 mL of methanol. The mixture was refluxed for 16 h, followed by theaddition of a second portion of N-Boc-3-bromopropanamine (7 g, 30 mmol).The mixture was refluxed for an additional 16 h, concentrated in vacuo,and purified by flash column chromatography (eluant, 10% MeOH/CH₂Cl₂).The product was dissolved in dichloromethane (50 mL), thentrifluoroacetic acid (50 mL) was added. After stirring at roomtemperature for 30 minutes, the solution was concentrated in vacuo, andthe resulting residue was suspended in ether (200 mL). The solidifiedresidue was collected by filtration, to yield the title intermediate asa TFA salt, which was converted to a neutral form by dissolving the saltin dichloromethane, then washing with aqueous NaOH solution.

b. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{8-(1S,3R,5R)-[3-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide

Potassium hydroxide (3 mg) dissolved in water (0.11 mL) and2,5-dihydrothiophene-1,1-dioxide (0.236 g, 2 mmol) was added to asolution of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[8-(1S,3R,5R)-(3-aminopropyl)-8-azabicyclo-[3.2.1]oct-3-yl]amide (freebase; 0.16 g, 0.4 mmol) in DMF (1 mL). The mixture was stirred at 75° C.for 16 h under nitrogen atmosphere. Evaporation in vacuo yielded thetitle intermediate, which was used in the next step without furtherpurification.

c. Synthesis of(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-(3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propyl)-carbamicacid methyl ester

N,N′-diisopropylethylamine (0.28 mL, 1.6 mmol) and methyl chloroformate(75 mg, 0.8 mmol) was added to a solution of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{8-(1S,3R,5R)-[3-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-ylamino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}-amide(0.206 g, 0.4 mmol) dissolved in DMF (2 mL). The reaction mixture wasstirred at room temperature for about 30 minutes, then concentrated invacuo. The residue was dissolved in 50% aqueous acetic acid (1 mL), andpurified by preparative HPLC to yield the title compound. (m/z): [M+H]⁺calcd for C₂₉H₄₀N₄O₆S 573.27; found, 573.6.

Example 9 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[2-(4-methanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)-amide

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid[(1S,3R,5R)-8-(2-ethenesulfonylethyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

Vinylsulfone (1.1 g, 9.32 mmol) was added dropwise to a stirred solutionof dichloromethane (25 mL) containing1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide (1.58 g, 4.64 mmol), theproduct of Example 1, step (f). The reaction mixture was stirred at roomtemperature overnight, then concentrated in vacuo, to yield the titleintermediate as an oily residue which was used in the next step withoutfurther purification.

b. Synthesis of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[2-(4-methanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

1-methylsulfonylpiperazine (656 mg, 0.4 mmol) was added to a solution of1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid[(1S,3R,5R)-8-(2-ethenesulfonyl-ethyl)-8-aza-bicyclo[3.2.1]oct-3-yl]-amide(45 mg, 0.1 mmol) in 1 mL of dichloromethane. The reaction mixture wasshaken at room temperature overnight, and concentrated in vacuo,yielding an oily residue. The residue was dissolved in 50% aqueousacetic acid (1 mL), then purified by preparative HPLC, to yield thetitle compound. (m/z): [M+H]⁺ calcd for C₂₉H₄₃N₅O₆S₂ 622.27; found,622.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.35 min.

Example 10 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(2-{2-[4-(tetrahydrofuran-2-carbonyl)-piperazin-1-yl]ethanesulfonyl}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

The title compound was prepared using the method described in Example 9by replacing, in Example 9, Step (b), 1-methylsulfonylpiperazine withpiperazin-1-yl-(tetra-hydrofuran-2-yl)methanone. (m/z): [M+H]⁺ calcd forC₃₃H₄₇N₅O₆S 642.32; found, 642.2. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.30 min.

Example 11 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[2-(4-ethanesulfonylpiperazin-1-yl)-ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)-amide

The title compound was prepared using the method described in Example 9by replacing in Example 9, Step (b) 1-methylsulfonylpiperazine with1-ethylsulfonyl-piperazine. (m/z): [M+H]⁺ calcd for C₃₀H₄₅N₅O₆S₂ 636.28;found, 636.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.41 min.

Example 12 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-acetylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

a. Preparation of1-{4-(3-chloropropane-1-sulfonyl)piperazin-1-yl}ethanone

N,N′-diisopropylethylamine (0.10 mL, 6 mmol) and then3-chloropropyl-1-sulfonyl chloride (53.1 mg, 0.3 mmol) were added to a 5mL glass vial containing N-acetylpiperazine (38 mg, 0.3 mmol) dissolvedin dichloromethane (1 mL). The reaction mixture was shaken at roomtemperature for about 0.5 h, then evaporated in vacuo, to yield thetitle intermediate as an oily residue which was used without furthertreatment.

b. Synthesis of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-acetylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

Sodium iodide (14 mg), N,N′-diisopropylethylamine (0.05 mL, 0.3 mmol),and 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide (45.3 mg, 0.1 mmol) wereadded to the product of the previous step dissolved in DMF (1 mL). Themixture was shaken at 85° C. for 24 h, then concentrated in vacuo. Theconcentrated residue was dissolved in 50% aqueous acetic acid (1 mL),then purified by preparative HPLC to yield the title compound. (m/z):[M+H]⁺ calcd for C₂₉H₄₁N₅O₅S 572.28; found 572.2. Retention time (anal.HPLC: 10-40% MeCN/H₂O over 6 min)=1.66 min.

Example 13 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[4-(tetrahydrofuran-2-carbonyl)piperazine-1-sulfonyl]propyl}-8-azabicyclo-[3.2.1]oct-3-yl)amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine withpiperazin-1-yl-(tetrahydro-furan-2-yl)-methanone. (m/z): [M+H]⁺ calcdfor C₃₂H₄₅N₅O₆S 628.31; found, 628.2. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=1.69 min.

Example 14 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-methanesulfonyl-piperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine with1-methylsulfonylpiperazine. (m/z): [M+H]⁺ calcd for C₂₈H₄₁N₅O₆S₂ 607.25;found, 608.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=1.61 min.

Example 15 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(3-{[1-(2-cyanoethyl)piperidin-4-yl]methylsulfamoyl}propyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine with3-(4-methylaminopiperidin-1-yl)propanenitrile. (m/z): [M+H]⁺ calcd forC₃₂H₄₆N₆O₄S 611.33; found, 611.20. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.32 min.

Example 16 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[(1-methanesulfonylpiperidin-4-yl)-methylsulfamoyl]-propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine with(1-methanesulfonylpiperidin-4-yl)methylamine. (m/z): [M+H]⁺ calcd forC₃₀H₄₅N₅O₆S₂ 636.28; found, 636.20. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=2.57 min.

Example 17 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-methylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine with1-methylpiperazine. (m/z): [M+H]⁺ calcd for C₂₈H₄₁N₅O₄S 544.29; found,544.3. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.21 min.

Example 18 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]propyl}-8-azabicyclo[3.2.1]oct-3-yl)amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine with2-piperazin-1-ylethanol. (m/z): [M+H]⁺ calcd for C₂₉H₄₃N₅O₅S 574.30;found, 574.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=1.19 min.

Example 19 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(3-dimethylaminopyrrolidine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

The title compound was prepared using the method described in Example 12by replacing in Example 12, Step (a), N-acetylpiperazine withdimethylpyrrolidin-3-ylamine. (m/z): [M+H]⁺ calcd for C₂₉H₄₃N₅O₄S558.30; found, 558.3. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.20 min.

Example 20 Synthesis of 4-acetylpiperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo-[3.2.1]oct-8-yl}propylester

a. Preparation of 4-acetylpiperazine-1-carboxylic acid 3-chloropropylester

N,N′-diisopropylethylamine (0.10 mL, 6 mmol), followed by3-chloropropane chloroformate (47.1 mg, 0.3 mmol) was added to a 5 mLglass vial containing 1-piperazin-1-yl-ethanone (38 mg, 0.3 mmol)dissolved in dichloromethane (1 mL). The reaction mixture was shaken atroom temperature for about 0.5 h, then evaporated in vacuo to yield thetitle intermediate as an oily residue which was used without furthertreatment.

b. Synthesis of 4-acetylpiperazine-1-carboxylic acid3-{(1S,3R5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo-[3.2.1]oct-8-yl}propylester

Sodium iodide (14 mg), followed by N,N′-diisopropylethylamine (0.05 mL,0.3 mmol), and 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-azabicyclo[3.2.1]oct-3-yl}amide (45.3 mg, 0.1 mmol) wereadded to the product of the previous step dissolved in DMF (1 mL). Themixture was shaken at 85° C. for 24 h, then concentrated in vacuo. Theresidue was dissolved in 50% aqueous acetic acid (1 mL), then purifiedby preparative HPLC to yield the title compound. (m/z): [M+H]⁺ calcd forC₃₀H₄₁N₅O₅ 552.31; found, 552.4. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=1.56 min.

Example 21 Synthesis of4-(tetrahydrofuran-2-carbonyl)piperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propylester

The title compound was prepared using the method described in Example 20by replacing, in Example 20, Step (a), 1-piperazin-1-ylethanone withpiperazin-1-yl-(tetrahydrofuran-2-yl)-methanone. (m/z): [M+H]⁺ calcd forC₃₃H₄₅N₅O₆ 607.34; found, 608.4. Retention time (anal. HPLC: 10-40%MeCN/H₂O over 6 min)=1.7 min.

Example 22 Synthesis of 4-methanesulfonylpiperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propylester

The title compound was prepared using the method described in Example 20by replacing in Example 20, Step (a), 1-piperazin-1-yl-ethanone with1-methylsulfonyl-piperazine. (m/z): [M+H]⁺ calcd for C₂₉H₄₁N₅O₆S 588.28;found, 588.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=1.74 min.

Example 23 Synthesis of 4-hydroxypiperidine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}propylester

The title compound was prepared using the method described in Example 20by replacing in Example 20, Step (a), 1-piperazin-1-yl-ethanone with4-hydroxy-piperidine. (m/z): [M+H]⁺ calcd for C₂₈H₃₉N₅O₆ 526.30; found,525.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.64 min.

Example 24 Synthesis of[2-(4-acetyl-piperazin-1-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo-[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester

a. Preparation of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid{(1S,3R,5R)-8-[2-(2,2-dimethoxyethylamino)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide

2,2-Dimethoxy-1-ethylamine (4.2 mL, 39 mmol) andN,N-diisopropylethylamine (4.53 mL, 26 mmol) were added to a solution of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[(2,2-dihydroxy)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amidehydrochloride salt (5.43 g, 13.0 mmol) in 30 mL of dichloromethane.After the mixture was stirred at room temperature for about 45 minutes,sodium triacetoxy-borohydride (3.86 g; 18.2 mmol) was added. The mixturewas stirred for about 4 h, then the remaining reducing agent wasquenched by adding water (20 mL) slowly to the reaction mixture in anice bath. The mixture was diluted with 200 mL of dichloromethane, andshaken in a funnel before collecting the organic layer. The organiclayer was washed with brine (50 mL) and a saturated sodium bicarbonatesolution, dried over MgSO₄, and evaporated to yield the titleintermediate which was used in the next step without further treatment.(m/z): [M+H]⁺ calcd for C₂₆H₃₈N₄O₄ 471.29; found, 472.0.

b. Preparation of(2,2-dimethoxyethyl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)amino]}-8-azabicyclo[3.2.1]oct-8-ylethyl)-carbamic acid methyl ester

Methyl chloroformate (0.275 mL, 3.58 mmol) andN,N′-diisopropylethylamine (0.62 mL, 3.58 mmol) were added to a coldsolution of 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[2-(2,2-dimethoxyethylamino)ethyl]-8-azabicyclo[3.2.1]oct-3-yl}amide(1.53 g, 3.25 mmol) dissolved in dichloromethane (25 mL) in an ice bath.The mixture was stirred at 0° C. for 2 h, then stirred at roomtemperature overnight. The mixture was diluted with dichloromethane (200mL), and washed with brine and a saturated sodium carbonate solution.After drying over MgSO₄, the organic solution was evaporated in vacuo,to yield an oily residue that was dissolved in 50% aqueous acetonitrile,then purified by preparative HPLC to yield the title intermediate.(m/z): [M+H]⁺ calcd for C₂₈H₄₀N₄O₆ 529.29; found, 529.3.

c. Preparation of(2,2-dihydroxy-ethyl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)-amino]-8-aza-bicyclo[3.2.1]oct-8-yl}-ethyl)-carbamicacid methyl ester

A solution of(2,2-dimethoxyethyl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester (236 mg, 0.367 mmol) in 6M HCl (5 mL) was stirred atroom temperature overnight. It was lyophilized to yield the titleintermediate as a hydrochloride salt.

d. Synthesis of[2-(4-acetyl-piperazin-1-yl)ethyl]-(2-{1(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo-[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester

1-piperazin-1-ylethanone (25.6 mg, 0.2 mmol),N,N′-diisopropyl-ethylamine (0.07 mL, 0.4 mmol), and sodiumtriacetoxyborohydride (29.7 mg, 0.14 mmol) were added to a solution of(2,2-dihydroxyethyl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)carbamicacid methyl ester (52 mg, 0.1 mmol) in 2 mL of dichloromethane. Themixture was shaken at room temperature for 2 h, then concentrated invacuo, yielding an oily residue. The residue was dissolved in 50%aqueous acetic acid (1 mL), purified by preparative HPLC, to yield thetitle compound. (m/z): [M+H]⁺ calcd for C₃₂H₄₆N₆O₅ 595.35; found, 595.2.Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.02 min

Example 25 Synthesis of(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carbonyl)-amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-[2-(4-methanesulfonylpiperazin-1-ylethyl]-carbamicacid methyl ester

The title compound was prepared using the method described in Example 24by replacing in Example 24, Step (d), 1-piperazin-1-ylethanone with1-methylsulfonyl-piperazine. (m/z): [M+H]⁺ calcd for C₃₁H₄₆N₆O₆S 631.32;found, 631.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.11 min.

Example 26 Synthesis of[2-(4-dimethylcarbamoylpiperazin-1-yl)-ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}-ethyl)-carbamicacid methyl ester

The title compound was prepared using the method described in Example 24by replacing in Example 24, Step (d), 1-piperazin-1-ylethanone withpiperazine-1-carboxylic acid dimethylamide. (m/z): [M+H]⁺ calcd forC₃₃H₄₉N₇O₅S 624.38; found, 624.3.

Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.07 min.

Example 27 Synthesis of[2-(1,1-dioxo-1λ⁶-thiomorpholin-4-yl)-ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]oct-8-yl}ethyl)-carbamicacid methyl ester

The title compound was prepared using the method described in Example 24by replacing in Example 24, Step (d), 1-piperazin-1-ylethanone withthiomorpholine-1,1-dioxide. (m/z): [M+H]⁺ calcd for C₃₀H₄₃N₅O₆S 602.29;found, 602.2. Retention time (anal. HPLC: 10-40% MeCN/H₂O over 6min)=2.16 min.

Example 28 Synthesis of1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(2-{[2-(4-dimethylcarbamoylpiperazin-1-yl)-ethyl]methanesulfonylamino}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]amide

The title compound was prepared using the method described in Example 24substituting the appropriate reagents. In Example 24, Step (b), methylchloroformate was replaced with methylsulfonyl chloride. In Example 24,Step (d), 1-piperazin-1-ylethanone was replaced withpiperazine-1-carboxylic acid dimethylamide to yield the title compound.(m/z): [M+H]⁺ calcd for C₃₂H₄₉N₇O₅S 644.35; found, 644.4. Retention time(anal. HPLC: 10-40% MeCN/H₂O over 6 min)=2.77 min.

Using the methods described in Examples 1-28, and substituting theappropriate reagents, the following compounds listed in Tables 1-5 wereprepared. In all of the compounds of the invention, thequinolinone-carboxamide is endo to the azabicyclooctanyl group.

TABLE 1 (I-c)

Calc'd Obsd No. a b X Q W Mol. Formula [M + H] [M + H] 1 1 2 N S(O)₂NS(O)₂CH₃ C₂₈H₄₁N₅O₆S₂ 608.25 608.2 2 1 2 C S(O)₂N(CH₃) NC(O)OCH₂CH₃C₃₀H₄₃N₅O₆S 602.31 602.2 3 1 2 C N{S(O)₂CH₃} NCH₃ C₃₀H₄₅N₅O₄S 572.34572.3 4 1 2 N S(O)₂ N(CH₂)₂OH C₂₉H₄₃N₅O₅S 574.30 574.2 5 1 2 N S(O)₂NCH₂-pyridin-4-yl C₃₃H₄₄N₆O₄S 621.33 621.3 6 1 2 N S(O)₂NCH₂-tetrahydro- C₃₂H₄₇N₅O₅S 614.35 614.3 furan-2-yl 7 1 2 N S(O)₂NCH₂C(O)N(CH₃)₂ C₃₁H₄₆N₆O₅S 615.34 615.3 8 1 2 N S(O)₂ N(CH₂)₃CH₃C₃₁H₄₇N₅O₄S 586.35 586.3 9 1 2 C S(O)₂NH NH C₂₈H₄₁N₅O₄S 544.30 544.3 101 2 N S(O)₂ NCH₃ C₂₈H₄₁N₅O₄S 544.29 544.3 11 0 2 C N{S(O)₂CH₃}N(CH₂)₂CH₃ C₃₁H₄₇N₅O₄S 586.33 586.3 12 0 2 C N{S(O)₂CH₃} N(CH₂)₂OHC₃₀H₄₅N₅O₅S 588.33 588.3 13 0 2 C N{S(O)₂CH₃} N(CH₂)₂OCH₃ C₃₁H₄₇N₅O₅S602.35 600.3 14 0 2 C N{S(O)₂CH₃} N(CH₂)₂-pyrrol-1-yl C₃₄H₄₈N₆O₄S 637.36637.3 15 0 2 C N{S(O)₂CH₃} NCH₂-pyridin-3-yl C₃₄H₄₆N₆O₄S 635.35 635.3 160 2 C N{S(O)₂CH₃} N(CH₂)₂NHC(O)OCH₃ C₃₂H₄₈N₆O₆S 645.35 645.3 17 0 2 CN{S(O)₂CH₃} N(CH₂)₂OC(O)N(CH₃)₂ C₃₃H₅₀N₆O₆S 659.37 659.3 18 0 2 CN{S(O)₂CH₃} N(CH₂)₂C(O)NHCH₃ C₃₃H₅₀N₆O₅S 643.37 643.3 19 0 2 CN{S(O)₂CH₃} N(CH₂)₃C(O)N(CH₃)₂ C₃₄H₅₂N₆O₅S 657.39 657.3 20 0 2 CN{S(O)₂CH₃} N(CH₂)₂NHS(O)₂CH₃ C₃₁H₄₈N₆O₆S₂ 665.32 665.3 21 0 2 CN{S(O)₂CH₃} N(CH₂)₂N(CH₃)S(O)₂ C₃₂H₅₀N₆O₆S₂ 679.34 679.3 —CH₃ 22 0 2 CN{S(O)₂CH₃} N(CH₂)₂S(O)₂N(CH₃)₂ C₃₃H₅₂N₆O₆S₂ 693.36 693.3 23 1 2 CS(O)₂N(CH₃) N(CH₂)₂CH₃ C₃₂H₄₉N₅O₄S 600.37 600.3 24 1 2 C S(O)₂N(CH₃)N(CH₂)₂OH C₃₁H₄₇N₅O₅S 602.35 602.3 25 1 2 C S(O)₂N(CH₃) N(CH₂)₂OCH₃C₃₂H₄₉N₅O₅S 616.35 616.2 26 1 2 C S(O)₂N(CH₃) N(CH₂)₂-pyrrol-1-ylC₃₅H₅₀N₆O₄S 651.38 651.3 27 1 2 C S(O)₂N(CH₃) NCH₂-pyridin-3-ylC₃₅H₄₈N₆O₄S 649.36 649.3 28 1 2 C S(O)₂N(CH₃) N(CH₂)₂OC(O)N(CH₃)₂C₃₄H₅₂N₆O₆S 673.38 673.3 29 1 2 C S(O)₂N(CH₃) N(CH₂)₂OC(O)NH—C₃₄H₅₂N₆O₅S 657.39 657.3 CH₂CH₃ 30 1 2 C S(O)₂N(CH₃) N(CH₂)₃C(O)N(CH₃)₂C₃₅H₅₄N₆O₅S 671.41 671.4 31 1 2 C S(O)₂N(CH₃) N(CH₂)₂NHS(O)₂CH₃C₃₂H₅₀N₆O₆S₂ 679.32 679.3 32 1 2 C S(O)₂N(CH₃) N(CH₂)₂N(CH₃)S(O)₂C₃₃H₅₂N₆O₆S₂ 693.36 693.3 —CH₃ 33 1 2 C S(O)₂N(CH₃) N(CH₂)₃S(O)₂N(CH₃)₂C₃₄H₅₄N₆O₆S₂ 707.37 707.3 34 1 2 C S(O)₂N(CH₃) NS(O)₂CH₃ C₃₀H₄₅N₅O₆S₂636.28 636.2 35 0 2 N S(O)₂ NS(O)₂CH₃ C₂₇H₃₉N₅O₆S₂ 594.25 594.2 36 0 2 NS(O)₂ NC(O)OCH₂CH₃ C₂₉H₄₁N₅O₆S 588.29 588.2 37 0 2 N S(O)₂ O C₂₆H₃₆N₄O₅S517.26 517.2 38 0 2 N S(O)₂ NCH₂-tetrahydro- C₃₁H₄₅N₅O₅S 600.33 600.3furan-2-yl 39 0 2 N S(O)₂ NCH₂C(O)N(CH₃)₂ C₃₀H₄₄N₆O₅S 601.33 601.3 40 02 N S(O)₂ NC(O)-tetrahydro- C₃₁H₄₃N₅O₆S 614.31 614.3 furan-2-yl 41 0 2 NS(O)₂ N-pyridin-4-yl C₃₁H₄₀N₆O₄S 593.30 593.2 42 0 2 N S(O)₂NCH₂-pyridin-4-yl C₃₂H₄₂N₆O₄S 607.31 607.2 43 0 2 N S(O)₂ N(CH₂)₂OHC₂₈H₄₁N₅O₅S 560.30 560.2 44 0 2 N S(O)₂ NCH₃ C₂₇H₃₉N₅O₄S 530.29 530.2 450 2 C N{S(O)₂CH₃} NC(O)OCH₃ C₃₀H₄₃N₅O₆S 602.31 602.2 46 0 2 CN{S(O)₂CH₃} NC(O)CH₃ C₃₀H₄₃N₅O₅S 586.31 586.3 47 0 2 C N{S(O)₂CH₃}NS(O)₂CF₃ C₂₉H₄₀F₃N₅O₆S₂ 676.25 676.2 48 0 2 C N{S(O)₂CH₃} NCH₂CF₃C₃₀H₄₂F₃N₅O₄S 626.31 626.2 49 0 2 C N{S(O)₂CH₃} N(CH₂)₂C(O)NH₂C₃₁H₄₆N₆O₅S 615.34 615.3 50 0 2 C N{S(O)₂CH₃} N(CH₂)₂CF₃ C₃₁H₄₄F₃N₅O₄S640.32 640.2 51 0 2 C N{S(O)₂CH₃} N(CH₂)₂CN C₃₁H₄₄N₆O₄S 597.33 597.2 520 2 C N{S(O)₂CH₃} N(CH₂)₂S(O)₂N(CH₃)₂ C₃₂H₅₀N₆O₆S₂ 679.34 679.2 53 0 2 CN{S(O)₂CH₃} NCH₂C(O)NH₂ C₃₀H₄₄N₆O₅S 601.33 601.2 54 0 2 C N{S(O)₂CH₃}NCH₂C(O)N(CH₃)₂ C₃₂H₄₈N₆O₅S 629.36 629.3 55 1 2 C S(O)₂N(CH₃)NCH₂C(O)N(CH₃)₂ C₃₃H₅₀N₆O₅S 643.37 643.2 56 1 2 C S(O)₂N(CH₃)NCH₂C(O)NH₂ C₃₁H₄₆N₆O₅S 615.34 615.2 57 1 2 C S(O)₂N(CH₃) NCH₂CF₃C₃₁H₄₄F₃N₅O₄S 640.32 640.2 58 1 2 C S(O)₂N(CH₃) N(CH₂)₂CF₃ C₃₂H₄₆F₃N₅O₄S654.34 654.2 59 1 3 C S(O)₂N(CH₃) N(CH₂)₂C(O)NH₂ C₃₂H₄₈N₆O₅S 629.36629.2 60 1 2 C S(O)₂N(CH₃) N(CH₂)₂CN C₃₂H₄₆N₆O₄S 611.33 611.2 61 1 2 CS(O)₂N(CH₃) NC(O)CH₂OCH₃ C₃₂H₄₇N₅O₆S 630.34 630.2 62 1 2 C S(O)₂N(CH₃)NCH₂CN C₃₁H₄₄N₆O₄S 597.33 597.2 63 0 1 C N{S(O)₂CH₃} S(O)₂ C₂₇H₃₈N₄O₆S₂579.22 579.2 64 0 1 C N{C(O)CH₃} S(O)₂ C₂₈H₃₈N₄O₅S 543.27 543.2 65 0 1 CN{C(O)N(CH₃)₂} S(O)₂ C₂₉H₄₁N₅O₅S 572.28 572.2 66 0 1 C N{C(O)OCH₃} S(O)₂C₂₈H₃₈N₄O₆S 559.25 559.2 67 0 1 C N{C(O)-pyridin-4-yl} S(O)₂ C₃₂H₃₉N₅O₅S606.28 606.2 68 0 2 N N{S(O)₂CH₃}(CH₂)₂ NS(O)₂CH₂CF₃ C₃₁H₄₅F₃N₆O₆S₂719.30 719.2 69 0 2 N N{S(O)₂CH₃}(CH₂)₂ S(O)₂ C₂₉H₄₃N₅O₆S₂ 622.28 622.270 0 2 N N{S(O)₂CH₃}(CH₂)₂ NC(O)N(CH₃)₂ C₃₂H₄₉N₇O₅S 644.35 644.4 71 0 2N N{S(O)₂CH₃}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₁H₄₉N₇O₆S₂ 680.34 680.2 72 0 2 NN{S(O)₂CH₃}(CH₂)₂ NC(O)NHCH₃ C₃₁H₄₇N₇O₅S 630.35 630.4 73 0 2 NS(O)₂(CH₂)₂ NS(O)₂N(CH₃)₂ C₂₈H₄₁N₅O₅S 560.30 560.2 74 0 1 CS(O)₂(CH₂)₂N(CH₃) NC(O)N(CH₃)₂ C₃₂H₄₈N₆O₅S 629.36 629.2 75 0 2 NS(O)₂(CH₂)₂ S(O)₂ C₂₈H₄₀N₄O₆S₂ 593.25 593.2 76 0 1 C S(O)₂(CH₂)₂N(CH₃)S(O)₂ C₂₉H₄₂N₄O₆S₂ 607.27 607.2 77 0 2 N S(O)₂(CH₂)₂ NS(O)₂CH₃C₂₉H₄₃N₅O₆S₂ 622.27 622.2 78 0 2 N S(O)₂(CH₂)₂ NC(O)-tetrahydro-C₃₃H₄₇N₅O₆S 642.32 642.2 furan-2-yl 79 0 2 N S(O)₂(CH₂)₂ NS(O)₂CH₂CF₃C₃₀H₄₂F₃N₅O₆S₂ 690.27 690.2 80 0 2 N S(O)₂(CH₂)₂ NS(O)₂CH₂CH₃C₃₀H₄₅N₅O₆S₂ 636.28 636.2 81 0 2 N S(O)₂(CH₂)₂ NS(O)₂CH(CH₃)₂C₃₁H₄₇N₅O₆S₂ 650.31 650.2 82 0 2 N S(O)₂(CH₂)₂ NC(O)OCH₃ C₃₀H₄₃N₅O₆S602.31 602.2 83 0 2 N S(O)₂(CH₂)₂ NC(O)N(CH₃)₂ C₃₁H₄₆N₆O₅S 615.34 615.284 0 3 N S(O)₂(CH₂)₂ NC(O)CH₃ C₃₁H₄₅N₅O₅S 600.33 600.2 85 0 1 CS(O)₂(CH₂)₂N(CH₃) NC(O)OCH₃ C₃₁H₄₅N₅O₆S 616.33 616.2 86 0 1 CS(O)₂(CH₂)₂N(CH₃) NS(O)₂CH₃ C₃₀H₄₅N₅O₆S₂ 636.30 636.2 87 0 2 CN{S(O)₂CH₃} S(O)₂ C₂₈H₄₀N₄O₆S₂ 593.25 593.1 88 0 2 C N{C(O)CH₃} S(O)₂C₂₉H₄₀N₄O₅S 557.29 557.2 89 0 2 C N{C(O)OCH₃} S(O)₂ C₂₉H₄₀N₄O₆S 573.27573.2 90 0 2 C N{C(O)-pyridin-4-yl} S(O)₂ C₃₃H₄₁N₅O₅S 620.30 620.2 91 01 C N{C(O)H} S(O)₂ C₂₇H₃₆N₄O₅S 529.26 529.2 92 0 1 C N{C(O)NHCH₃} S(O)₂C₂₈H₃₉N₅O₅S 558.27 558.2 93 0 1 C N{C(O)NH₂} S(O)₂ C₂₇H₃₇N₅O₅S 544.27544.2 94 0 2 N N{S(O)₂N(CH₃)₂}(CH₂)₂ NS(O)₂CH₃ C₃₁H₄₉N₇O₆S₂ 680.34 680.295 0 2 N N{S(O)₂N(CH₃)₂}(CH₂)₂ NC(O)N(CH₃)₂ C₃₃H₅₂N₈O₅S 673.40 673.2 960 2 N N{S(O)₂N(CH₃)₂}(CH₂)₂ NC(O)OCH₃ C₃₂H₄₉N₇O₆S 660.36 660.2 97 0 2 NN{S(O)₂N(CH₃)₂}(CH₂)₂ NC(O)CH₃ C₃₂H₄₉N₇O₅S 644.37 644.2 98 0 2 NN{S(O)₂N(CH₃)₂}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₂H₅₂N₈O₆S₂ 709.36 709.2 99 0 2 NN{S(O)₂N(CH₃)₂}(CH₂)₂ S(O)₂ C₃₀H₄₆N₆O₆S₂ 651.31 651.2 100 0 2 NN{C(O)OCH₃)(CH₂)₂ NS(O)₂CH₃ C₃₁H₄₆N₆O₆S 631.32 631.2 101 0 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)N(CH₃)₂ C₃₃H₄₉N₇O₅ 624.38 624.3 102 0 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)OCH₃ C₃₂H₄₆N₆O₆ 611.31 611.2 103 0 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)CH₃ C₃₂H₄₆N₆O₅ 595.35 595.2 104 0 2 NN{C(O)OCH₃}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₂H₄₉N₇O₆S 660.36 660.2 105 0 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)-tetrahydro- C₃₅H₅₀N₆O₆ 651.40 651.3 furan-2-yl106 0 2 N N{C(O)OCH₃}(CH₂)₂ S(O)₂ C₃₀H₄₃N₅O₆S 602.29 602.2 107 0 1 CN{C(O)OCH₃}— S(O)₂ C₃₁H₄₅N₅O₆S 616.33 616.2 (CH₂)₂N(CH₃) 108 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NS(O)₂CH₃ C₃₂H₄₉N₇O₅S 644.37 644.2 109 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NC(O)N(CH₃)₂ C₃₄H₅₂N₈O₄ 637.43 637.3 110 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NC(O)OCH₃ C₃₃H₄₉N₇O₅ 624.40 624.3 111 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NC(O)CH₃ C₃₃H₄₉N₇O₄ 608.40 608.3 112 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₃H₅₂N₈O₅S 673.40 673.2 113 0 2 NN{C(O)N(CH₃)₂}(CH₂)₂ NC(O)-tetrahydro- C₃₆H₅₃N₇O₅ 664.43 664.3furan-2-yl 114 0 1 C N{C(O)N(CH₃)₂}— S(O)₂ C₃₂H₄₈N₆O₅S 629.36 629.2(CH₂)₂N(CH₃) 115 0 2 C N{C(O)H} S(O)₂ C₂₈H₃₈N₄O₅S 543.27 543.1 116 0 2 CN{C(O)NHCH₃} S(O)₂ C₂₉H₄₁N₅O₅S 572.30 572.2 117 0 2 C N{C(O)NH₂} S(O)₂C₂₈H₃₉N₅O₅S 558.28 558.1 118 1 1 C N{C(O)OCH₃} S(O)₂ C₂₉H₄₀N₄O₆S 573.27573.6 119 1 2 N N{C(O)OCH₃}(CH₂)₂ NS(O)₂CH₃ C₃₂H₄₈N₆O₆S 645.35 645.4 1201 2 N N{C(O)OCH₃}(CH₂)₂ NC(O)N(CH₃)₂ C₃₄H₅₁N₇O₅ 638.41 638.4 121 1 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)OCH₃ C₃₃H₄₈N₆O₆ 625.38 625.4 122 1 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)CH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 123 1 2 NN{C(O)OCH₃}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₆S 674.38 674.4 124 1 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)-tetrahydro- C₃₆H₅₂N₆O₆ 665.41 665.4 furan-2-yl125 1 3 N N{C(O)OCH₃}(CH₂)₂ NS(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 126 1 3 NN{C(O)OCH₃}(CH₂)₂ NC(O)CH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 127 1 2 NN{C(O)OCH₃}(CH₂)₂ NC(O)NHCH₃ C₃₃H₄₉N₇O₅ 624.40 624.4 128 1 2 NN{C(O)OCH₃}(CH₂)₂ S(O)₂ C₃₁H₄₅N₅O₆S 616.33 616.4 129 1 2 C N{C(O)OCH₃}—S(O)₂ C₃₃H₄₉N₅O₆S 644.36 644.4 (CH₂)₂N(CH₃) 130 1 1 C N{C(O)OCH₃}— S(O)₂C₃₂H₄₇N₅O₆S 630.34 630.4 (CH₂)₂N(CH₃) 131 1 1 C N{C(O)OCH₃}—NS(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.40 688.4 (CH₂)₂N(CH₃) 132 1 1 CN{C(O)OCH₃}— NC(O)OCH₃ C₃₄H₅₀N₆O₆ 639.40 639.4 (CH₂)₂N(CH₃) 133 1 1 CN{C(O)OCH₃}— NS(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 (CH₂)₂N(CH₃) 134 1 2 NN{C(O)CH₃}(CH₂)₂ NS(O)₂CH₃ C₃₂H₄₈N₆O₅S 629.36 629.4 135 1 2 NN{C(O)CH₃}(CH₂)₂ NC(O)N(CH₃)₂ C₃₄H₅₁N₇O₄ 622.42 622.4 136 1 2 NN{C(O)CH₃}(CH₂)₂ NC(O)OCH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 137 1 2 NN{C(O)CH₃}(CH₂)₂ NC(O)CH₃ C₃₃H₄₈N₆O₄ 593.39 593.4 138 1 2 NN{C(O)CH₃}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₅S 658.38 658.4 139 1 2 NN{C(O)CH₃}(CH₂)₂ NC(O)-tetrahydro- C₃₆H₅₂N₆O₅ 649.42 649.4 furan-2-yl140 1 2 N N{C(O)CH₃}(CH₂)₂ NS(O)₂CH₂S(O)₂— C₃₃H₅₀N₆O₇S₂ 707.34 707.2 CH₃141 1 3 N N{C(O)CH₃}(CH₂)₂ NS(O)₂CH₃ C₃₃H₅₀N₆O₅S 643.37 643.4 142 1 3 NN{C(O)CH₃}(CH₂)₂ NC(O)CH₃ C₃₄H₅₀N₆O₄ 607.41 607.4 143 1 2 NN{C(O)CH₃}(CH₂)₂ NC(O)NHCH₃ C₃₃H₄₉N₇O₄ 608.40 608.4 144 1 2 NN{C(O)CH₃}(CH₂)₂ S(O)₂ C₃₁H₄₅N₅O₅S 600.33 600.4 145 1 2 CN{C(O)CH₃}(CH₂)₂ S(O)₂ C₃₃H₄₉N₅O₅S 628.36 628.4 146 1 1 CN{C(O)CH₃}(CH₂)₂ S(O)₂ C₃₂H₄₇N₅O₅S 614.35 614.4 —N(CH₃) 147 1 1 CN{C(O)CH₃}(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₅S 672.40 672.4 —N(CH₃) 148 1 1C N{C(O)CH₃}(CH₂)₂ NC(O)OCH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 —N(CH₃) 149 1 1 CN{C(O)CH₃}(CH₂)₂ NS(O)₂CH₃ C₃₃H₅₀N₆O₅S 643.37 643.4 —N(CH₃) 150 1 2 NS(O)₂N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₁H₄₈N₆O₆S₂ 665.32 665.2 151 1 2 NS(O)₂N(CH₃)(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₅S 658.38 658.3 152 1 2 NS(O)₂N(CH₃)(CH₂)₂ NC(O)OCH₃ C₃₂H₄₈N₆O₆S 645.35 645.3 153 1 2 NS(O)₂N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₂H₄₈N₆O₅S 629.36 629.3 154 1 2 NS(O)₂N(CH₃)(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₂H₅₁N₇O₆S₂ 694.35 694.3 155 1 3 NS(O)₂N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₃H₅₀N₆O₅S 643.37 643.3 156 1 2 NS(O)₂N(CH₃)(CH₂)₂ NC(O)NHCH₃ C₃₂H₄₉N₇O₅S 644.37 644.3 157 1 1 CS(O)₂N(CH₃)(CH₂)₂— NS(O)₂N(CH₃)₂ C₃₃H₅₃N₇O₆S₂ 708.37 708.3 N(CH₃) 158 12 N S(O)₂N(CH₃)(CH₂)₂ NC(O)-tetrahydro- C₃₅H₅₂N₆O₆S 685.37 685.3furan-2-yl 159 1 2 N S(O)₂N(CH₃)(CH₂)₂ NS(O)₂CH₂S(O)₂CH₃ C₃₂H₅₀N₆O₈S₃743.30 743.2 160 1 3 N S(O)₂N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₂H₅₀N₆O₆S₂ 679.34679.3 161 1 2 C S(O)₂N(CH₃)(CH₂)₂— S(O)₂ C₃₂H₄₉N₅O₆S₂ 664.33 664.3N(CH₃) 162 1 1 C S(O)₂N(CH₃)(CH₂)₂— S(O)₂ C₃₁H₄₇N₅O₆S₂ 650.31 650.3N(CH₃) 163 1 1 C S(O)₂N(CH₃)(CH₂)₂— NC(O)OCH₃ C₃₃H₅₀N₆O₆S 659.37 659.3N(CH₃) 164 0 2 C N{S(O)₂CH₃} NC(O)N(CH₃)₂ C₃₁H₄₆N₆O₅S 615.34 615.4 165 02 C N{S(O)₂CH₃} NC(O)NHCH₃ C₃₀H₄₄N₆O₅S 601.33 601.2 166 0 2 CN{C(O)N(CH₃)₂} NC(O)OCH₃ C₃₂H₄₆N₆O₅ 595.37 595.4 167 0 2 CN{C(O)N(CH₃)₂} NC(O)CH₃ C₃₂H₄₆N₆O₄ 579.37 579.4 168 0 2 C N{C(O)N(CH₃)₂}NC(O)N(CH₃)₂ C₃₃H₄₉N₇O₄ 608.40 608.4 169 0 2 C N{C(O)N(CH₃)₂ NC(O)NHCH₃C₃₂H₄₇N₇O₄ 594.39 594.4 170 0 2 C N{C(O)N(CH₃)₂} NS(O)₂CH₃ C₃₁H₄₆N₆O₅S615.34 615.4 171 0 2 C N{S(O)₂CH₃} NC(O)NH₂ C₂₉H₄₂N₆O₅S 587.31 587.2 1720 2 C N{C(O)N(CH₃)₂} NC(O)NH₂ C₃₁H₄₅N₇O₄ 580.37 580.2 173 1 2 NS(O)₂N(CH₃)(CH₂)₂ S(O)₂ C₃₀H₄₅N₅O₆S₂ 636.30 636.2 174 1 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 175 1 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)N(CH₃)₂ C₃₄H₅₁N₇O₅ 638.41 638.4 176 1 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)OCH₃ C₃₃H₄₈N₆O₆ 625.38 625.4 177 1 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)tetrahydro- C₃₆H₅₂N₆O₆ 665.41 665.4 furan-2-yl178 1 2 N OC(O)N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₂H₄₈N₆O₆S 645.35 645.4 179 1 2 NOC(O)N(CH₃)(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₆S 674.38 674.4 180 1 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)NHCH₃ C₃₃H₄₉N₇O₅ 624.40 624.4 181 1 3 NOC(O)N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 182 1 3 NOC(O)N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 183 1 2 NOC(O)N(CH₃)(CH₂)₂ S(O)₂ C₃₁H₄₅N₅O₆S 616.33 616.4 184 1 2 COC(O)N(CH₃)(CH₂)₂— S(O)₂ C₃₃H₄₉N₅O₆S 644.36 643.9 N(CH₃) 185 1 1 COC(O)N(CH₃)(CH₂)₂— S(O)₂ C₃₂H₄₇N₅O₆S 630.34 630.4 N(CH₃) 186 1 1 COC(O)N(CH₃)(CH₂)₂— NS(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.40 688.4 N(CH₃) 187 0 2N OC(O)N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₂H₄₆N₆O₅ 595.37 595.4 188 0 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)N(CH₃)₂ C₃₃H₄₉N₇O₅ 624.40 624.4 189 0 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)OCH₃ C₃₂H₄₆N₆O₆ 611.36 611.4 190 0 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)-tetrahydro- C₃₅H₅₀N₆O₆ 651.40 651.4 furan-2-yl191 0 2 N OC(O)N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₁H₄₆N₆O₆S 631.34 631.4 192 0 2 NOC(O)N(CH₃)(CH₂)₂ NS(O)₂N(CH₃)₂ C₃₂H₄₉N₇O₆S 660.36 660.4 193 0 2 NOC(O)N(CH₃)(CH₂)₂ NC(O)NHCH₃ C₃₂H₄₇N₇O₅ 610.38 610.4 194 0 3 NOC(O)N(CH₃)(CH₂)₂ NC(O)CH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 195 0 3 NOC(O)N(CH₃)(CH₂)₂ NS(O)₂CH₃ C₃₂H₄₈N₆O₆S 645.35 645.4 196 0 2 NOC(O)N(CH₃)(CH₂)₂ S(O)₂ C₃₀H₄₃N₅O₆S 602.31 602.2 197 0 1 C OC(O)N(CH₃)—S(O)₂ C₃₁H₄₅N₅O₆S 616.33 616.2 (CH₂)₂N(CH₃) 198 0 1 C OC(O)N(CH₃)—NS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₆S 674.38 674.4 (CH₂)₂N(CH₃) 199 1 2 N SCH₂C(O)NC(O)CH₃ C₃₁H₄₃N₅O₄S 582.32 582.2 200 1 2 N SCH₂C(O) NC(O)N(CH₃)₂C₃₂H₄₆N₆O₄S 611.35 611.4 201 1 2 N S(O)₂(CH₂)₂ NC(O)OCH₃ C₃₁H₄₅N₅O₆S616.33 616.2 202 1 2 N S(O)₂(CH₂)₂ NC(O)CH₃ C₃₁H₄₅N₅O₅S 600.31 600.4 2031 2 N S(O)₂(CH₂)₂ NS(O)₂CH₃ C₃₀H₄₅N₅O₆S₂ 636.30 636.2 204 1 2 NS(O)₂(CH₂)₂ NC(O)N(CH₃)₂ C₃₂H₄₈N₆O₅S 629.36 629.4 205 1 2 N S(O)₂(CH₂)₂S(O)₂ C₂₉H₄₂N₄O₆S₂ 607.27 607.2 206 1 2 N S(O)₂CH₂C(O) NC(O)CH₃C₃₁H₄₃N₅O₆S 614.31 614.2 207 1 1 C S(O)₂CH₂C(O)N(CH₃) S(O)₂ C₃₀H₄₂N₄O₇S₂635.26 635.2 208 1 2 N S(O)₂CH₂C(O) NC(O)OCH₃ C₃₁H₄₃N₅O₇S 630.30 630.2209 1 2 N S(O)₂CH₂C(O) NC(O)-tetrahydro- C₃₄H₄₇N₅O₇S 670.34 670.2furan-2-yl 210 1 2 N S(O)₂CH₂C(O) S(O)₂ C₂₉H₄₀N₄O₇S₂ 621.25 621.2 211 11 C S(O)₂CH₂C(O)N(CH₃) NS(O)₂N(CH₃)₂ C₃₂H₄₈N₆O₇S₂ 693.32 693.2 212 1 2 NS(O)₂ NC(O)tetrahydro- C₃₂H₄₅N₅O₆S 628.31 628.2 furan-2-yl 213 1 2 NS(O)₂ NC(O)CH₃ C₂₉H₄₁N₅O₅S 572.28 572.2 214 1 1 C S(O)₂N(CH₃) S(O)₂C₂₈H₄₀N₄O₆S₂ 593.25 593.2 215 1 2 N S(O)₂ S(O)₂ C₂₇H₃₈N₄O₆S₂ 579.24579.2 216 1 1 C S(O)₂N(CH₃) NS(O)₂N(CH₃)₂ C₃₀H₄₆N₆O₆S₂ 651.31 651.2 2171 2 N S(O)₂ NC(O)OCH₃ C₂₉H₄₁N₅O₆S 588.29 588.2 218 1 2 N OC(O) NS(O)₂CH₃C₂₉H₄₁N₅O₆S 588.28 588.2 219 1 2 N OC(O) NC(O)tetrahydro- C₃₃H₄₅N₅O₆608.34 608.4 furan-2-yl 220 1 1 C OC(O)N(CH₃) S(O)₂ C₂₉H₄₀N₄O₆S 573.28573.2 221 1 1 C OC(O)N(CH₃) NS(O)₂N(CH₃)₂ C₃₁H₄₆N₆O₆S 631.34 631.2 222 12 N OC(O) NC(O)OCH₃ C₃₀H₄₁N₅O₆ 568.32 568.2 223 1 2 N OC(O) NC(O)CH₃C₃₀H₄₁N₅O₅ 552.31 552.4 224 1 2 N

NC(O)OCH₃ C₃₃H₄₈N₆O₆S 657.35 657.4 225 1 2 N

NC(O)tetrahydro- furan-2-yl C₃₆H₅₂N₆O₆S 697.38 697.4 226 1 2 N

NC(O)CH₃ C₃₃H₄₈N₆O₅S 641.36 641.4 227 1 2 N

NS(O)₂CH₃ C₃₂H₄₈N₆O₆S₂ 677.32 677.2 228 1 3 N

NC(O)CH₃ C₃₄H₅₀N₆O₅S 655.37 655.4 229 1 3 N

NS(O)₂CH₃ C₃₃H₅₀N₆O₆S₂ 691.34 691.4 230 1 2 N

NC(O)tetrahydro- furan-2-yl C₃₇H₅₂N₆O₆ 677.41 677.4 231 1 2 N

NC(O)CH₃ C₃₄H₄₈N₆O₅ 621.39 621.4 232 1 2 N

NS(O)₂CH₃ C₃₃H₄₈N₆O₆S 657.35 657.4 233 1 3 N

NC(O)CH₃ C₃₅H₅₀N₆O₅ 635.40 635.4 234 1 3 N

NS(O)₂CH₃ C₃₄H₅₀N₆O₆S 671.37 671.4

TABLE 2 (I-d)

Molecular Calc'd Obsd No. a b Q Y Formula [M + H] [M + H] 235 1 1 S(O)₂3-N(CH₃)₂ C₂₉H₄₃N₅O₄S 558.30 558.3 236 1 2 S(O)₂ 4-CH₂NHS(O)₂CH₃C₃₀H₄₅N₅O₆S₂ 636.30 636.3 237 0 1 S(O)₂ 3-N(CH₃)C(O)CH₃ C₂₉H₄₁N₅O₅S572.30 572.2 238 0 1 S(O)₂ 3-N(CH₃)₂ C₂₈H₄₁N₅O₄S 544.30 544.2 239 0 2S(O)₂ 4-OH C₂₇H₃₈N₄O₅S 531.27 531.1 240 0 2 N{S(O)₂CH₃}(CH₂)₂3-OC(O)N(CH₃)₂ C₃₃H₅₀N₆O₆S 659.37 659.4 241 0 2 N{S(O)₂CH₃}(CH₂)₂3-NHC(O)CH₃ C₃₂H₄₈N₆O₅S 629.36 629.4 242 0 2 N{S(O)₂CH₃}(CH₂)₂ 2-C(O)NH₂C₃₁H₄₆N₆O₅S 615.34 615.2 243 0 2 N{S(O)₂CH₃}(CH₂)₂ 3-N(CH₃)S(O)₂CH₃C₃₂H₅₀N₆O₆S₂ 679.34 679.2 244 0 2 S(O)₂(CH₂)₂ 3-N(CH₃)C(O)OCH₃C₃₂H₄₇N₅O₆S 630.34 630.2 245 0 2 S(O)₂(CH₂)₂ 3-N(CH₃)C(O)N(CH₃)₂C₃₃H₅₀N₆O₅S 643.37 643.4 246 0 2 S(O)₂(CH₂)₂ 3-NHC(O)CH₃ C₃₁H₄₅N₅O₅S600.33 600.2 247 0 2 S(O)₂(CH₂)₂ 4-CH₂NHS(O)₂CH₃ C₃₁H₄₇N₅O₆S₂ 650.31650.2 248 0 2 S(O)₂(CH₂)₂ 3-OC(O)N(CH₃)₂ C₃₂H₄₇N₅O₆S 630.34 630.2 249 12 N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)S(O)₂CH₃ C₃₄H₅₂N₆O₆S 673.38 673.4 250 1 2N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)C(O)OCH₃ C₃₅H₅₂N₆O₆ 653.41 653.4 251 1 2N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)C(O)CH₃ C₃₅H₅₂N₆O₅ 637.42 637.4 252 1 2N{C(O)OCH₃}(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.40 688.4 253 1 2N{C(O)OCH₃}(CH₂)₂ 3-NHC(O)OCH₃ C₃₄H₅₀N₆O₆ 639.40 639.4 254 1 2N{C(O)OCH₃}(CH₂)₂ 3-NHC(O)CH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 255 1 2N{C(O)OCH₃}(CH₂)₂ 3-NHC(O)N(CH₃)₂ C₃₅H₅₃N₇O₅ 652.43 652.4 256 1 2N{C(O)OCH₃}(CH₂)₂ 3-C(O)NH₂ C₃₃H₄₈N₆O₅ 609.39 609.4 257 1 2N{C(O)OCH₃}(CH₂)₂ 4-OC(O)N(CH₃)₂ C₃₅H₅₂N₆O₆ 653.41 653.4 258 1 2N{C(O)OCH₃}(CH₂)₂ 4-CH₂NHS(O)₂CH₃ C₃₄H₅₂N₆O₆S 673.38 673.4 259 1 1N{C(O)OCH₃}(CH₂)₂ 3-NHC(O)CH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 260 1 1N{C(O)OCH₃}(CH₂)₂ 3-NHC(O)OCH₃ C₃₃H₄₈N₆O₆ 625.38 625.4 261 1 1N{C(O)OCH₃}(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₆S 674.38 674.4 262 1 1N{C(O)OCH₃}(CH₂)₂ 3-OC(O)N(CH₃)₂ C₃₄H₅₀N₆O₆ 639.40 639.4 263 1 1N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)C(O)CH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 264 1 1N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)S(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.40 688.4 265 1 1N{C(O)OCH₃}(CH₂)₂ 3-N(CH₃)C(O)N(CH₃)₂ C₃₅H₅₃N₇O₅ 652.43 652.4 266 1 2N{C(O)CH₃}(CH₂)₂ 3-N(CH₃)S(O)₂CH₃ C₃₄H₅₂N₆O₅S 657.39 657.4 267 1 2N{C(O)CH₃}(CH₂)₂ 3-N(CH₃)C(O)OCH₃ C₃₅H₅₂N₆O₅ 637.42 637.4 268 1 2N{C(O)CH₃}(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₅S 672.40 672.4 269 1 2N{C(O)CH₃}(CH₂)₂ 3-NHC(O)OCH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 270 1 2N{C(O)CH₃}(CH₂)₂ 3-NHC(O)CH₃ C₃₄H₅₀N₆O₄ 607.41 607.4 271 1 2N{C(O)CH₃}(CH₂)₂ 3-NHC(O)N(CH₃)₂ C₃₅H₅₃N₇O₄ 636.43 636.4 272 1 2N{C(O)CH₃}(CH₂)₂ 3-C(O)NH₂ C₃₃H₄₈N₆O₄ 593.39 593.4 273 1 2N{C(O)CH₃}(CH₂)₂ 4-OC(O)N(CH₃)₂ C₃₅H₅₂N₆O₅ 637.42 637.4 274 1 2N{C(O)CH₃}(CH₂)₂ 4-CH₂NHS(O)₂CH₃ C₃₄H₅₂N₆O₅S 657.39 657.4 275 1 1N{C(O)CH₃}(CH₂)₂ 3-NHC(O)CH₃ C₃₃H₄₈N₆O₄ 593.39 593.4 276 1 1N{C(O)CH₃}(CH₂)₂ 3-NHC(O)OCH₃ C₃₃H₄₈N₆O₅ 609.39 609.4 277 1 1N{C(O)CH₃}(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₃H₅₁N₇O₅S 658.38 658.4 278 1 1N{C(O)CH₃}(CH₂)₂ 3-OC(O)N(CH₃)₂ C₃₄H₅₀N₆O₅ 623.40 623.4 279 1 1N{C(O)CH₃}(CH₂)₂ 3-N(CH₃)C(O)CH₃ C₃₄H₅₀N₆O₄ 607.41 607.4 280 1 1N{C(O)CH₃}(CH₂)₂ 3-N(CH₃)S(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₅S 672.40 672.4 281 1 1N{C(O)CH₃}(CH₂)₂ 3-N(CH₃)C(O)N(CH₃)₂ C₃₅H₅₃N₇O₄ 636.43 636.4 282 1 2S(O)₂N(CH₃)(CH₂)₂ 3-N(CH₃)S(O)₂CH₃ C₃₃H₅₂N₆O₆S₂ 693.36 693.3 283 1 2S(O)₂N(CH₃)(CH₂)₂ 3-N(CH₃)C(O)OCH₃ C₃₄H₅₂N₆O₆S 673.38 673.3 284 1 2S(O)₂N(CH₃)(CH₂)₂ 3-N(CH₃)C(O)CH₃ C₃₄H₅₂N₆O₅S 657.39 657.3 285 1 2S(O)₂N(CH₃)(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₃H₅₃N₇O₆S₂ 708.37 708.3 286 1 2S(O)₂N(CH₃)(CH₂)₂ 3-NHC(O)OCH₃ C₃₃H₅₀N₆O₆S 659.37 659.3 287 1 2S(O)₂N(CH₃)(CH₂)₂ 3-NHC(O)CH₃ C₃₃H₅₀N₆O₅S 643.37 643.3 288 1 2S(O)₂N(CH₃)(CH₂)₂ 3-NHC(O)N(CH₃)₂ C₃₄H₅₃N₇O₅S 672.40 672.3 289 1 2S(O)₂N(CH₃)(CH₂)₂ 3-C(O)NH₂ C₃₂H₄₈N₆O₅S 629.36 629.3 290 1 2S(O)₂N(CH₃)(CH₂)₂ 4-CH₂NHS(O)₂CH₃ C₃₃H₅₂N₆O₆S₂ 693.36 693.2 291 1 1S(O)₂N(CH₃)(CH₂)₂ 3-NHC(O)CH₃ C₃₂H₄₈N₆O₅S 629.36 629.3 292 1 1S(O)₂N(CH₃)(CH₂)₂ 3-NHC(O)OCH₃ C₃₂H₄₈N₆O₆S 645.35 645.3 293 1 1S(O)₂N(CH₃)(CH₂)₂ 3-NHS(O)₂N(CH₃)₂ C₃₂H₅₁N₇O₆S₂ 694.35 694.3 294 1 1S(O)₂N(CH₃)(CH₂)₂ 3-N(CH₃)C(O)CH₃ C₃₃H₅₀N₆O₅S 643.37 643.3 295 1 1S(O)₂N(CH₃)(CH₂)₂ 3-N(CH₃)S(O)₂N(CH₃)₂ C₃₃H₅₃N₇O₆S₂ 708.37 708.3 296 1 2S(O)₂N(CH₃)(CH₂)₂ 3-OC(O)N(CH₃)₂ C₃₄H₅₂N₆O₆S 673.38 673.4 297 1 2S(O)₂N(CH₃)(CH₂)₂ 3-OC(O)N(CH₃)₂ C₃₀H₄₅N₅O₆S₂ 636.30 636.2 298 1 2OC(O)N(CH₃)(CH₂)₂ 3-C(O)NH₂ C₃₃H₄₈N₆O₅ 609.39 609.4 299 1 2OC(O)N(CH₃)(CH₂)₂ 3-N(CH₃)S(O)₂CH₃ C₃₄H₅₂N₆O₆S 673.38 673.4 300 1 2OC(O)N(CH₃)(CH₂)₂ 4-CH₂NHS(O)₂CH₃ C₃₄H₅₂N₆O₆S 673.38 673.4 301 1 2OC(O)N(CH₃)(CH₂)₂ 4-OH C₃₂H₄₇N₅O₅ 582.37 582.4 302 1 2 OC(O)N(CH₃)(CH₂)₂4-OC(O)N(CH₃)₂ C₃₅H₅₂N₆O₆ 653.41 653.4 303 1 1 OC(O)N(CH₃)(CH₂)₂3-OC(O)N(CH₃)₂ C₃₄H₅₀N₆O₆ 639.40 639.4 304 1 1 OC(O)N(CH₃)(CH₂)₂3-N(CH₃)S(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 305 1 1 OC(O)N(CH₃)(CH₂)₂3-N(CH₃)S(O)₂N(CH₃)₂ C₃₄H₅₃N₇O₆S 688.40 688.4 306 1 1 OC(O)N(CH₃)(CH₂)₂3-N(CH₃)C(O)CH₃ C₃₄H₅₀N₆O₅ 623.40 623.4 307 0 2 OC(O)N(CH₃)(CH₂)₂3-C(O)NH₂ C₃₂H₄₆N₆O₅ 595.37 595.4 308 0 2 OC(O)N(CH₃)(CH₂)₂3-N(CH₃)S(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 309 0 2 OC(O)N(CH₃)(CH₂)₂4-CH₂NHS(O)₂CH₃ C₃₃H₅₀N₆O₆S 659.37 659.4 310 0 2 OC(O)N(CH₃)(CH₂)₂ 4-OHC₃₁H₄₅N₅O₅ 568.36 568.4 311 0 2 OC(O)N(CH₃)(CH₂)₂ 4-OC(O)N(CH₃)₂C₃₄H₅₀N₆O₆ 639.40 639.4 312 0 1 OC(O)N(CH₃)(CH₂)₂ 3-OC(O)N(CH₃)₂C₃₃H₄₈N₆O₆ 625.38 625.4 313 0 1 OC(O)N(CH₃)(CH₂)₂ 3-N(CH₃)S(O)₂CH₃C₃₂H₄₈N₆O₆S 645.35 645.4 314 0 1 OC(O)N(CH₃)(CH₂)₂ 3-N(CH₃)S(O)₂N(CH₃)₂C₃₃H₅₁N₇O₆S 674.38 674.4 315 0 1 OC(O)N(CH₃)(CH₂)₂ 3-N(CH₃)C(O)CH₃C₃₃H₄₈N₆O₅ 609.39 609.4 316 1 2 SCH₂C(O) 4-N(CH₃)S(O)₂CH₃ C₃₂H₄₇N₅O₅S₂646.32 646.2 317 1 2 S(O)₂(CH₂)₂ 4-N(CH₃)S(O)₂CH₃ C₃₂H₄₉N₅O₆S₂ 664.33664.2 318 1 2 S(O)₂CH₂C(O) 4-N(CH₃)S(O)₂CH₃ C₃₂H₄₇N₅O₇S₂ 678.31 678.2319 1 2 S(O)₂CH₂C(O) 3-C(O)NH₂ C₃₁H₄₃N₅O₆S 614.31 614.2 320 1 2 S(O)₂4-CH₂NHC(O)OCH₃ C₃₁H₄₅N₅O₆S 616.33 616.2 321 1 2 S(O)₂4-CH₂N(CH₃)S(O)₂CH₃ C₃₁H₄₇N₅O₆S₂ 650.31 650.2 322 1 2 S(O)₂ 4-OHC₂₈H₄₀N₄O₅S 545.29 545.2 323 1 2 S(O)₂ 4-OC(O)N(CH₃)₂ C₃₁H₄₅N₅O₆S 616.33616.2 324 1 2 S(O)₂ 3-C(O)NH₂ C₂₉H₄₁N₅O₅S 572.30 572.2 325 1 2 S(O)₂3-N(CH₃)S(O)₂CH₃ C₃₀H₄₅N₅O₆S₂ 636.30 636.2 326 1 1 S(O)₂ 3-N(CH₃)C(O)CH₃C₃₀H₄₃N₅O₅S 586.31 586.2 327 1 1 S(O)₂ 3-OC(O)N(CH₃)₂ C₃₀H₄₃N₅O₆S 602.31602.2 328 1 2 OC(O) 4-CH₂NHC(O)OCH₃ C₃₂H₄₅N₅O₆ 596.35 596.4 329 1 2OC(O) 4-CH₂NHS(O)₂CH₃ C₃₁H₄₅N₅O₆S 616.33 616.2 330 1 2 OC(O)4-CH₂N(CH₃)S(O)₂CH₃ C₃₂H₄₇N₅O₆S 630.34 630.4 331 1 2 OC(O)4-OC(O)N(CH₃)₂ C₃₂H₄₅N₅O₆ 596.35 596.4 332 1 2 OC(O) 3-C(O)NH₂C₃₀H₄₁N₅O₅ 552.33 552.2 333 1 2 OC(O) 3-N(CH₃)C(O)CH₃ C₃₂H₄₅N₅O₅ 580.36580.4 334 1 2 OC(O) 3-N(CH₃)S(O)₂CH₃ C₃₁H₄₅N₅O₆S 616.33 616.2 335 1 1OC(O) 3-N(CH₃)C(O)CH₃ C₃₁H₄₃N₅O₅ 566.34 566.4 336 1 1 OC(O)3-OC(O)N(CH₃)₂ C₃₁H₄₃N₅O₆ 582.34 582.4 337 1 2 OC(O) 3-CH₂OH C₃₀H₄₂N₄O₅539.33 539.4 338 1 2 OC(O) 4-OH C₂₉H₄₀N₄O₅ 525.30 525.2 339 1 2

4-OH C₃₂H₄₇N₅O₅S 614.35 614.2 340 1 2

4-CH₂NHS(O)₂CH₃ C₃₄H₅₂N₆O₆S₂ 705.36 705.4 341 1 2

3-C(O)NH₂ C₃₃H₄₈N₆O₅S 641.36 641.4 342 1 1

3-N(CH₃)S(O)₂CH₃ C₃₃H₅₀N₆O₆S₂ 691.34 691.2 343 1 1

3-N(CH₃)CH₂— C(O)N(CH₃)₂ C₃₂H₄₈N₆O₅S 629.36 629.4 344 1 2

4-OC(O)N(CH₃)₂ C₃₅H₅₂N₆O₆S 685.38 685.4 345 1 2

4-OH C₃₃H₄₇N₅O₅ 594.37 594.4 346 1 2

4-OC(O)N(CH₃)₂ C₃₆H₅₂N₆O₆ 665.41 665.4 347 1 2

4-CH₂NHS(O)₂CH₃ C₃₅H₅₂N₆O₆S 685.38 685.4 348 1 2

4-CH₂NHC(O)OCH₃ C₃₆H₅₂N₆O₆ 665.41 665.4 349 1 1

3-N(CH₃)S(O)₂CH₃ C₃₄H₅₀N₆O₆S 671.37 671.4 350 1 1

3-OC(O)N(CH₃)₂ C₃₅H₅₀N₆O₆ 651.40 651.4 351 1 1

3-N(CH₃)CH₂— C(O)N(CH₃)₂ C₃₃H₄₈N₆O₅ 609.39 609.4

TABLE 3 (I)

Molecular Calc'd Obsd No. a Z Formula [M + H] [M + H] 352 0

C₂₉H₄₁N₅O₅S 572.30 572.3 353 1

C₂₈H₃₉N₅O₅S 558.28 558.3 354 0

C₂₇H₃₈N₄O₅S 531.27 531.2 355 0

C₃₁H₄₈N₆O₆S 633.35 633.2 356 0

C₃₁H₄₅N₅O₆ 584.35 584.2 357 0

C₃₁H₄₄N₆O₅ 581.35 581.2 358 0

C₃₂H₄₈N₆O₅ 597.39 597.3 359 0

C₃₂H₄₇N₇O₄ 594.39 594.3 360 0

C₂₇H₃₈N₄O₄S₂ 547.25 547.2 361 0

C₂₈H₃₈N₄O₄S 527.28 527.2 362 0

C₃₂H₃₉N₅O₃S 574.29 574.2 363 0

C₂₇H₃₆N₄O₃S 497.27 497.2 364 0

C₂₈H₃₉N₅O₃S 526.29 526.2 365 0

C₂₈H₃₈N₄O₃S 511.28 511.2 366 0

C₂₇H₃₇N₅O₃S 512.28 512.2 367 0

C₂₉H₄₁N₅O₃S 540.31 540.2 368 0

C₂₈H₃₉N₅O₃S 526.29 526.2 369 0

C₂₇H₃₇N₅O₃S 512.28 512.2 370 0

C₂₈H₃₈N₄O₃S 511.28 511.2 371 1

C₃₂H₄₇N₅O₆ 598.37 598.4 372 1

C₃₃H₅₀N₆O₆S 659.37 659.4 373 1

C₃₂H₄₆N₆O₅ 595.37 595.4 374 1

C₃₂H₄₇N₅O₅ 582.37 582.4 375 1

C₃₃H₅₀N₆O₅S 643.37 643.4 376 1

C₃₂H₄₆N₆O₄ 579.37 579.4 377 0

C₂₈H₃₈N₄O₆S 559.27 559.4 378 0

C₂₈H₃₈N₄O₆S 559.27 559.4 379 1

C₃₁H₄₆N₆O₅S 615.34 615.3 380 1

C₃₁H₄₇N₅O₆S 618.34 618.3 381 1

C₃₂H₅₀N₆O₆S₂ 679.34 679.3 382 1

C₃₁H₄₆N₆O₅S 615.34 615.2 383 1

C₃₃H₅₁N₇O₆S 674.38 674.4 384 1

C₃₂H₄₆N₆O₅ 595.37 595.4 385 1

C₃₂H₄₆N₆O₅ 595.37 595.4 386 0

C₃₂H₄₉N₇O₆S 660.36 660.4 387 0

C₃₃H₄₈N₆O₅ 609.39 609.4 388 0

C₃₁H₄₄N₆O₅ 581.35 581.4 389 0

C₃₁H₄₄N₆O₅ 581.35 581.4 390 1

C₃₀H₄₃N₅O₅S 586.31 586.2 391 1

C₃₀H₄₁N₅O₆S 600.29 600.2 392 1

C₃₀H₄₃N₅O₅S 586.31 586.2 393 1

C₃₀H₄₄N₆O₆S 617.32 617.2 394 1

C₃₂H₄₆N₆O₅S 627.34 627.4 395 1

C₃₃H₄₆N₆O₅ 607.37 607.4

TABLE 4 (I-c)

No. a b X Q W 396 0 2 C N{S(O)₂CH₃} NH 397 1 2 C S(O)₂N(CH₃) NH 398 0 2C N{C(O)N(CH₃)₂} S(O)₂ 399 0 1 C N{C(O)-pyridin-4-yl} S 400 0 2 CN{S(O)₂CH₃} NS(O)CH₃ 401 0 2 C N{C(O)CH₃} NC(O)CH₃ 402 0 2 C N{C(O)H}NC(O)H 403 0 1 C N{S(O)₂CH₃} NS(O)₂CH₃ 404 0 1 C N{C(O)CH₃} NC(O)CH₃ 4050 1 C N{C(O)H} NC(O)H 406 1 2 C N{C(O)CH₃} NC(O)CH₃ 407 1 2 C N{C(O)H}NC(O)H 408 1 1 C N{C(O)CH₃} NC(O)CH₃ 409 1 1 C N{C(O)H} NC(O)H 410 0 2 CN{C(O)N(CH₃)₂} NC(O)H 411 1 1 C S(O)₂N(CH₃)(CH₂)₂N(CH₃) NS(O)₂CH₃ 412 12 N SCH₂C(O) NS(O)₂CH₃ 413 1 2 N S(O)₂CH₂C(O) NS(O)₂CH₃ 414 1 2 N OC(O)N(CH₂)₂OH 415 1 2 N OC(O) S(O)₂

TABLE 5 (I)

No. a Z 416 0

417 0

418 0

419 0

420 0

421 0

422 1

423 1

Example 29 Radioligand Binding Assay on 5-HT_(4(c)) Human Receptors

a. Membrane Preparation 5-HT_(4(c))

HEK-293 (human embryonic kidney) cells stably-transfected with human5-HT_(4(c)) receptor cDNA (Bmax=˜6.0 μmol/mg protein, as determinedusing [³H]-GR113808 membrane radioligand binding assay) were grown inT-225 flasks in Dulbecco's Modified Eagles Medium (DMEM) containing4,500 mg/L D-glucose and pyridoxine hydrochloride (GIBCO-InvitrogenCorp., Carlsbad Calif.: Cat #11965) supplemented with 10% fetal bovineserum (FBS) (GIBCO-Invitrogen Corp.: Cat #10437), 2 mM L-glutamine and(100 units) penicillin-(100 μg) streptomycin/ml (GIBCO-Invitrogen Corp.:Cat #15140) in a 5% CO₂, humidified incubator at 37° C. Cells were grownunder continuous selection pressure by the addition of 800 μg/mLgeneticin (GIBCO-Invitrogen Corp.: Cat # 10131) to the medium.

Cells were grown to roughly 60-80% confluency (<35 subculture passages).At 20-22 hours prior to harvesting, cells were washed twice and fed withserum-free DMEM. All steps of the membrane preparation were performed onice. The cell monolayer was lifted by gentle mechanical agitation andtrituration with a 25 mL pipette. Cells were collected by centrifugationat 1000 rpm (5 min).

For the membrane preparation, cell pellets were resuspended in ice-cold50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES), pH7.4 (membrane preparation buffer) (40 mL/total cell yield from 30-40T225 flasks) and homogenized using a polytron disrupter (setting 19,2×10 s) on ice. The resultant homogenates were centrifuged at 1200 g for5 min at 4° C. The pellet was discarded and the supernatant centrifugedat 40,000 g (20 min). The pellet was washed once by resuspension withmembrane preparation buffer and centrifugation at 40,000 g (20 min). Thefinal pellet was resuspended in 50 mM HEPES, pH 7.4 (assay buffer)(equivalent 1 T225 flask/1 mL). Protein concentration of the membranesuspension was determined by the method of Bradford (Bradford, 1976).Membranes were stored frozen in aliquots at −80° C.

b. Radioligand Binding Assays

Radioligand binding assays were performed in 1.1 mL 96-deep wellpolypropylene assay plates (Axygen) in a total assay volume of 400 μLcontaining 2 μg membrane protein in 50 mM HEPES pH 7.4, containing0.025% bovine serum albumin (BSA). Saturation binding studies fordetermination of K_(d) values of the radioligand were performed using[³H]-GR113808 (Amersham Inc., Bucks, UK: Cat #TRK944; specific activity˜82 Ci/mmol) at 8-12 different concentrations ranging from 0.001 nM-5.0nM. Displacement assays for determination of pK_(i) values of compoundswere performed with [³H]-GR113808 at 0.15 nM and eleven differentconcentrations of compound ranging from 10 μM-100 μM.

Test compounds were received as 10 mM stock solutions in DMSO anddiluted to 400 μM into 50 mM HEPES pH 7.4 at 25° C., containing 0.1%BSA, and serial dilutions (1:5) then made in the same buffer.Non-specific binding was determined in the presence of 1 μM unlabeledGR113808. Assays were incubated for 60 min at room temperature, and thenthe binding reactions were terminated by rapid filtration over 96-wellGF/B glass fiber filter plates (Packard BioScience Co., Meriden, Conn.)presoaked in 0.3% polyethyleneimine. Filter plates were washed threetimes with filtration buffer (ice-cold 50 mM HEPES, pH7.4) to removeunbound radioactivity. Plates were dried, 35 μL Microscint-20 liquidscintillation fluid (Packard BioScience Co., Meriden, Conn.) was addedto each well and plates were counted in a Packard Topcount liquidscintillation counter (Packard BioScience Co., Meriden, Conn.).

Binding data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 3-parameter model for one-site competition. The BOTTOM(curve minimum) was fixed to the value for nonspecific binding, asdetermined in the presence of 1 μM GR113808. K_(i) values for testcompounds were calculated, in Prism, from the best-fit IC₅₀ values, andthe K_(d) value of the radioligand, using the Cheng-Prusoff equation(Cheng and Prusoff, Biochemical Pharmacology, 1973, 22, 3099-108):K_(i)=IC₅₀/(1+[L]/K_(d)) where [L]=concentration [³H]-GR113808. Resultsare expressed as the negative decadic logarithm of the K_(i) values,pK_(i).

Test compounds having a higher pK_(i) value in this assay have a higherbinding affinity for the 5-HT₄ receptor. The compounds of the inventionwhich were tested in this assay had a pK_(i) value ranging from about6.3 to about 9.4, typically ranging from about 6.5 to about 8.5.

Example 30 Radioligand Binding Assay on 5-HT_(3A) Human ReceptorsDetermination of Receptor Subtype Selectivity

a. Membrane Preparation 5-HT_(3A)

HEK-293 (human embryonic kidney) cells stably-transfected with human5-HT_(3A) receptor cDNA were obtained from Dr. Michael Bruess(University of Bonn, GDR) (Bmax=˜9.0 μmol/mg protein, as determinedusing [³H]-GR65630 membrane radioligand binding assay). Cells were grownin T-225 flasks or cell factories in 50% Dulbecco's Modified EaglesMedium (DMEM) (GIBCO-Invitrogen Corp., Carlsbad, Calif.: Cat #11965) and50% Ham's F12 (GIBCO-Invitrogen Corp.: Cat #11765) supplemented with 10%heat inactivated fetal bovine serum (FBS) (Hyclone, Logan, Utah: Cat#SH30070.03) and (50 units) penicillin-(50 μg) streptomycin/ml(GIBCO-Invitrogen Corp.: Cat #15140) in a 5% CO₂, humidified incubatorat 37° C.

Cells were grown to roughly 70-80% confluency (<35 subculture passages).All steps of the membrane preparation were performed on ice. To harvestthe cells, the media was aspirated and cells were rinsed with Ca²⁺,Mg²⁺-free Dulbecco's phosphate buffered saline (dPBS). The cellmonolayer was lifted by gentle mechanical agitation. Cells werecollected by centrifugation at 1000 rpm (5 min). Subsequent steps of themembrane preparation followed the protocol described above for themembranes expressing 5-HT₄(c) receptors.

b. Radioligand Binding Assays

Radioligand binding assays were performed in 96-well polypropylene assayplates in a total assay volume of 200 μL containing 1.5-2 μg membraneprotein in 50 mM HEPES pH 7.4, containing 0.025% BSA assay buffer.Saturation binding studies for determination of K_(d) values of theradioligand were performed using [³H]-GR65630 (PerkinElmer Life SciencesInc., Boston, Mass.: Cat #NET1011, specific activity ˜85 Ci/mmol) attwelve different concentrations ranging from 0.005 nM to 20 nM.Displacement assays for determination of pK_(i) values of compounds wereperformed with [³H]-GR65630 at 0.50 mM and eleven differentconcentrations of compound ranging from 10 pM to 100 μM. Compounds werereceived as 10 mM stock solutions in DMSO (see section 3.1), diluted to400 μM into 50 mM HEPES pH 7.4 at 25° C., containing 0.1% BSA, andserial (1:5) dilutions then made in the same buffer. Non-specificbinding was determined in the presence of 10 μM unlabeled MDL72222.Assays were incubated for 60 min at room temperature, then the bindingreactions were terminated by rapid filtration over 96-well GF/B glassfiber filter plates (Packard BioScience Co., Meriden, Conn.) presoakedin 0.3% polyethyleneimine. Filter plates were washed three times withfiltration buffer (ice-cold 50 mM HEPES, pH7.4) to remove unboundradioactivity. Plates were dried, 35 μL Microscint-20 liquidscintillation fluid (Packard BioScience Co., Meriden, Conn.) was addedto each well and plates were counted in a Packard Topcount liquidscintillation counter (Packard BioScience Co., Meriden, Conn.).

Binding data were analyzed using the non-linear regression proceduredescribed above to determine K_(i) values. The BOTTOM (curve minimum)was fixed to the value for nonspecific binding, as determined in thepresence of 10 μM MDL72222. The quantity [L] in the Cheng-Prusoffequation was defined as the concentration[³H]-GR65630.

Selectivity for the 5-HT₄ receptor subtype with respect to the 5-HT₃receptor subtype was calculated as the ratioK_(i)(5-HT_(3A))/K_(i)(5-HT₄(C)). The compounds of the invention whichwere tested in this assay had a 5-HT₄/5-HT₃ receptor subtype selectivityranging from about 10 to about 95,000, typically ranging from about 100to about 4000.

Example 31 Whole-cell cAMP Accumulation Flashplate Assay with HEK-293cells expressing human 5-HT₄(c) Receptors

In this assay, the functional potency of a test compound was determinedby measuring the amount of cyclic AMP produced when HEK-293 cellsexpressing 5-HT₄ receptors were contacted with different concentrationsof test compound.

a. Cell Culture

HEK-293 (human embryonic kidney) cells stably-transfected with clonedhuman 5-HT₄(c) receptor cDNA were prepared expressing the receptor attwo different densities: (1) at a density of about 0.5-0.6 μmol/mgprotein, as determined using a [³H]-GR113808 membrane radioligandbinding assay, and (2) at a density of about 6.0 μmol/mg protein. Thecells were grown in T-225 flasks in Dulbecco's Modified Eagles Medium(DMEM) containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat#11965) supplemented with 10% fetal bovine serum (FBS) (GIBCO-InvitrogenCorp.: Cat #10437) and (100 units) penicillin-(100 μg) streptomycin/ml(GIBCO-Invitrogen Corp.: Cat #15140) in a 5% CO₂, humidified incubatorat 37° C. Cells were grown under continuous selection pressure by theaddition of geneticin (800 μg/mL: GIBCO-Invitrogen Corp.: Cat #10131) tothe medium.

b. Cell Preparation

Cells were grown to roughly 60-80% confluency. Twenty to twenty-twohours prior to assay, cells were washed twice, and fed, with serum-freeDMEM containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat#11965). To harvest the cells, the media was aspirated and 10 mL Versene(GIBCO-Invitrogen Corp.: Cat #15040) was added to each T-225 flask.Cells were incubated for 5 min at RT and then dislodged from the flaskby mechanical agitation. The cell suspension was transferred to acentrifuge tube containing an equal volume of pre-warmed (37° C.) dPBSand centrifuged for 5 min at 1000 rpm. The supernatant was discarded andthe pellet was re-suspended in pre-warmed (37° C.) stimulation buffer(10 mL equivalent per 2-3 T-225 flasks). This time was noted and markedas time zero. The cells were counted with a Coulter counter (count above8 μm, flask yield was 1-2×10⁷ cells/flask). Cells were resuspended at aconcentration of 5×10⁵ cells/ml in pre-warmed (37° C.) stimulationbuffer (as provided in the flashplate kit) and preincubated at 37° C.for 10 min.

cAMP assays were performed in a radioimmunoassay format using theFlashplate Adenylyl Cyclase Activation Assay System with ¹²⁵I-cAMP(SMP004B, PerkinElmer Life Sciences Inc., Boston, Mass.), according tothe manufacturer's instructions.

Cells were grown and prepared as described above. Final cellconcentrations in the assay were 25×10³ cells/well and the final assayvolume was 100 μL. Test compounds were received as 10 mM stock solutionsin DMSO, diluted to 400 mM into 50 mM HEPES pH 7.4 at 25° C., containing0.1% BSA, and serial (1:5) dilutions then made in the same buffer.Cyclic AMP accumulation assays were performed with 11 differentconcentrations of compound ranging from 10 pM to 100 μM (final assayconcentrations). A 5-HT concentration-response curve (10 μM to 100 μM)was included on every plate. The cells were incubated, with shaking, at37° C. for 15 min and the reaction terminated by addition of 100 μl ofice-cold detection buffer (as provided in the flashplate kit) to eachwell. The plates were sealed and incubated at 4° C. overnight. Boundradioactivity was quantified by scintillation proximity spectroscopyusing the Topcount (Packard BioScience Co., Meriden, Conn.).

The amount of cAMP produced per mL of reaction was extrapolated from thecAMP standard curve, according to the instructions provided in themanufacturer's user manual. Data were analyzed by nonlinear regressionanalysis with the GraphPad Prism Software package using the 3-parametersigmoidal dose-response model (slope constrained to unity). Potency dataare reported as pEC₅₀ values, the negative decadic logarithm of the EC₅₀value, where EC₅₀ is the effective concentration for a 50% maximalresponse.

Test compounds exhibiting a higher pEC₅₀ value in this assay have ahigher potency for agonizing the 5-HT₄ receptor. The compounds of theinvention which were tested in this assay, for example, in the cell line(1) having a density of about 0.5-0.6 μmol/mg protein, had a pEC₅₀ valueranging from about 7.0 to about 9.5, typically ranging from about 7.5 toabout 8.5.

Example 32 In Vitro Voltage Clamp Assay of Inhibition of Potassium IonCurrent in Whole Cells Expressing the hERG Cardiac Potassium Channel

CHO-K¹ cells stably transfected with HERG cDNA were obtained from GailRobertson at the University of Wisconsin. Cells were held in cryogenicstorage until needed. Cells were expanded and passaged in Dulbecco'sModified Eagles Medium/F12 supplemented with 10% fetal bovine serum and200 μg/mL geneticin. Cells were seeded onto poly-D-lysine (100 μg/mL)coated glass coverslips, in 35 mm² dishes (containing 2 mL medium) at adensity that enabled isolated cells to be selected for whole cellvoltage-clamp studies. The dishes were maintained in a humidified, 5%CO₂ environment at 37° C.

Extracellular solution was prepared at least every 7 days and stored at4° C. when not in use. The extracellular solution contained (mM): NaCl(137), KCl (4), CaCl₂ (1.8), MgCl₂ (1), Glucose (10),4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES) (10), pH 7.4with NaOH. The extracellular solution, in the absence or presence oftest compound, was contained in reservoirs, from which it flowed intothe recording chamber at approximately 0.5 mL/min. The intracellularsolution was prepared, aliquoted and stored at −20° C. until the day ofuse. The intracellular solution contained (mM): KCl (130), MgCl₂ (1),ethylene glycol-bis(beta-aminoethyl ether) N,N,N′,N′-tetra acetic acidsalt (EGTA) (5), MgATP (5),4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES) (10), pH 7.2with KOH. All experiments were performed at room temperature (20-22°C.).

The coverslips on which the cells were seeded were transferred to arecording chamber and perfused continuously. Gigaohm seals were formedbetween the cell and the patch electrode. Once a stable patch wasachieved, recording commenced in the voltage clamp mode, with theinitial holding potential at −80 mV. After a stable whole-cell currentwas achieved, the cells were exposed to test compound. The standardvoltage protocol was: step from the holding potential of −80 mV to +20mV for 4.8 sec, repolarize to −50 mV for 5 sec and then return to theoriginal holding potential (−80 mV). This voltage protocol was run onceevery 15 sec (0.067 Hz). Peak current amplitudes during therepolarization phase were determined using pClamp software. Testcompounds at a concentration of 3 μM were perfused over the cells for 5minutes, followed by a 5-minute washout period in the absence ofcompound. Finally a positive control (cisapride, 20 nM) was added to theperfusate to test the function of the cell. The step from −80 mV to +20mV activates the hERG channel, resulting in an outward current. The stepback to −50 mV results in an outward tail current, as the channelrecovers from inactivation and deactivates.

Peak current amplitudes during the repolarization phase were determinedusing pCLAMP software. The control and test article data were exportedto Origin® (OriginLab Corp., Northampton Mass.) where the individualcurrent amplitudes were normalized to the initial current amplitude inthe absence of compound. The normalized current means and standarderrors for each condition were calculated and plotted versus the timecourse of the experiment.

Comparisons were made between the observed K⁺ current inhibitions afterthe five-minute exposure to either the test article or vehicle control(usually 0.3% DMSO). Statistical comparisons between experimental groupswere performed using a two-population, independent t-test (MicrocalOrigin v. 6.0). Differences were considered significant at p<0.05.

The smaller the percentage inhibition of the potassium ion current inthis assay, the smaller the potential for test compounds to change thepattern of cardiac repolarization when used as therapeutic agents. Thecompounds of the invention which were tested in this assay at aconcentration of 3 μM typically exhibited an inhibition of the potassiumion current of less than about 20%. For example, the compound of Example17 when tested in this assay exhibited an inhibition of the potassiumion current of less than about 15%.

Example 33 Pharmacokinetic Study in the Rat

Aqueous solution formulations of test compounds were prepared in 0.1%lactic acid at a pH of between about 5 and about 6. Male Sprague-Dawleyrats (CD strain, Charles River Laboratories, Wilmington, Mass.) weredosed with test compounds via intravenous administration (IV) at a doseof 2.5 mg/kg or by oral gavage (PO) at a dose of 5 mg/kg. The dosingvolume was 1 mL/kg for IV and 2 mL/kg for PO administration. Serialblood samples were collected from animals pre-dose, and at 2 (IV only),5, 15, and 30 min, and at 1, 2, 4, 8, and 24 hours post-dose.Concentrations of test compounds in blood plasma were determined byliquid chromatography-mass spectrometry analysis (LC-MS/MS) (MDS SCIEX,API 4000, Applied Biosystems, Foster City, Calif.) with a lower limit ofquantitation of 1 ng/mL.

Standard pharmacokinetic parameters were assessed by non-compartmentalanalysis (Model 201 for IV and Model 200 for PO) using WinNonlin(Version 4.0.1, Pharsight, Mountain View, Calif.). The maximum in thecurve of test compound concentration in blood plasma vs. time is denotedC_(max). The area under the concentration vs. time curve from the timeof dosing to the last measurable concentration (AUC(0-t)) was calculatedby the linear trapezoidal rule. Oral bioavailability (F(%)), i.e. thedose-normalized ratio of AUC(0-t) for PO administration to AUC(0-t) forIV administration, was calculated as:F(%)=AUC_(PO)/AUC_(IV)×Dose_(IV)/Dose_(PO)×100%

Test compounds which exhibit larger values of the parameters C_(max),AUC(0-t), and F(%) in this assay are expected to have greaterbioavailability when administered orally. The compounds of the inventionthat were tested in this assay typically had C_(max) values ranging fromabout 0.01 to about 1.2 μg/mL, more typically ranging from about 0.1 toabout 0.5 μg/mL, and AUC(0-t) values typically ranging from about 0.15to about 1.4 μg·hr/mL, more typically ranging from about 0.2 to about0.8 μg·hr/mL. By way of example, the compound of Example 17 when testedin this assay had a C_(max) value of 0.06 μg/mL, an AUC(0-t) value of0.45 μg·hr/mL and oral bioavailability (F(%)) in the rat model of about20%.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto. Additionally, all publications, patents, andpatent documents cited hereinabove are incorporated by reference hereinin full, as though individually incorporated by reference.

1. A method of treating a disorder of reduced motility of thegastrointestinal tract in a mammal, the method comprising administeringto the mammal a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of formula (I):

wherein: R¹ is hydrogen, halo, or C₁₋₄alkyl; R² is C₃₋₄alkyl orC₃₋₆cycloalkyl; a is 0 or 1; Z is a moiety of formula (c):

wherein: X is carbon and Q is -A-; or X is nitrogen and Q is selectedfrom —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, and -A(CH₂)₂—; b is 1, X iscarbon, and W is —S(O)₂—; or b is 2, X is carbon or nitrogen, and W isselected from —S(O)₂—, —N{C(O)R⁹}—, —N{S(O)₂R¹⁰}—, —N{C(O)NR¹³R¹⁴}—, and—N{R¹⁶}—; or Z is a moiety of formula (d):

wherein: Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—, and-A(CH₂)₂—; W′ is selected from —N(R⁸)C(O)R⁹, —N(R⁸)S(O)₂R¹⁰,—S(R¹¹)(O)₂, —N(R⁸)C(O)NR¹³R¹⁴, —OR¹⁵, and —N(R⁸)R¹⁶; provided that whenW′ is attached to a carbon atom bonded to the nitrogen atom of the ring,that W′ is —C(O)NR¹³R¹⁴; and b is 1 or 2; A is selected from—S(O)₂CH₂C(O)N(R³)—, —N{C(O)R⁵}—, —N{C(O)NR^(6a)R^(6b)}—,—N{S(O)₂C₁₋₃alkyl}—, —N{S(O)₂NR^(6a)R^(6b)}—, —S(O)₂N(R^(7a))—, and—OC(O)N(R^(7b))—; R³ is C₁₋₄alkyl; R⁵ is hydrogen, C₁₋₃alkyl, orC₁₋₃alkoxy; R^(6a) and R^(6b) are independently hydrogen or C₁₋₄alkyl;R^(7a) and R^(7b) are independently hydrogen or C₁₋₄alkyl; R⁸ ishydrogen, methyl, or ethyl; R⁹ is tetrahydrofuranyl, methyl, or ethyl;R¹⁰ is methyl or ethyl; R¹¹ is methyl or ethyl; R¹³ and R¹⁴ areindependently hydrogen, methyl or ethyl; R¹⁵ is hydrogen or methyl; R¹⁶is —(CH₂)_(r)—R¹⁷, wherein r is 0, 1, or 2; and R¹⁷ is selected fromhydroxy, cyano, C₁₋₃alkyl, and C₁₋₃alkoxy; provided that when r is 0,R¹⁷ is C₁₋₃alkyl; and when r is 1, R¹⁷ is cyano or C₁₋₃alkyl; or apharmaceutically-acceptable salt or stereoisomer thereof.
 2. The methodof claim 1, wherein Z is a moiety of formula (c), wherein X is nitrogenand Q is selected from —OC(O)—, —S(O)₂—, —S(O)₂(CH₂)₂—,—S(O)₂N(R^(7a))(CH₂)₂—, —N{C(O)C₁₋₃alkoxy}(CH₂)₂—, and—N{S(O)₂C₁₋₃alkyl}(CH₂)₂—.
 3. The method of claim 1, wherein Z is amoiety of formula (c), wherein X is carbon and Q is selected from—N{C(O)C₁₋₃alkoxy}-, —N{C(O)NR^(6a)R^(6b)}—, —N{S(O)₂C₁₋₃alkyl}, and—S(O)₂N(R^(7a))—.
 4. The method of claim 1, wherein Z is a moiety offormula (d), wherein Q is selected from —OC(O)— and —S(O)₂—.
 5. Themethod of claim 1, wherein the compound of formula (I) is selected from:1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-methanesulfonylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(3-dimethylaminopyrrolidine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-{3-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]propyl}-8-azabicyclo[3.2.1]-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-methylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[methanesulfonyl-(1-propylpiperidin-4-yl)amino]ethyl}-8-azabicyclo[3.2.1]-3-yl)-amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(3-{[1-(2-methoxyethyl)piperidin-4-yl]methylsulfamoyl]propyl)-8-azabicyclo[3.2.1]-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[(1-methanesulfonylpiperidin-4-yl)methylsulfamoyl]propyl}-8-azabicyclo[3.2.1]-3-yl]amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(3-{[1-(2-cyanoethyl)piperidin-4-yl]methylsulfamoyl}propyl)-8-azabicyclo[3.2.1]-3-yl]amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)methanesulfonylamino]ethyl}-8-azabicyclo[3.2.1]-3-yl)amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3,3-dimethylureido]ethyl}-8-azabicyclo[3.2.1]-oct-3-yl)amide;(1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-carbamicacid methyl ester; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid[(1S,3R,5R)-8-(2-{[2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]methanesulfonylamino}ethyl)-8-azabicyclo-[3.2.1]-3-yl]amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[2-(4-methanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid[(1S,3R,5R)-8-(2-{2-[4-(tetrahydrofuran-2-carbonyl)piperazin-1-yl]ethanesulfonyl}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{2-[2-(4-ethanesulfonylpiperazin-1-yl)ethanesulfonyl]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;(1,1-dioxo-hexahydro-1λ⁶-thiopyran-4-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-carbamicacid methyl ester; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylicacid((1S,3R,5R)-8-{2-[1-(1,1-dioxotetrahydro-1λ⁶-thiophen-3-yl)-3-methylureido]ethyl}-8-azabicyclo[3.2.1]oct-3-yl)amide;(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)-amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-[2-(4-methanesulfonylpiperazin-1-ylethyl]-carbamicacid methyl ester;[2-(4-dimethylcarbamoylpiperazin-1-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}-ethyl)-carbamicacid methyl ester;[2-(4-acetyl-piperazin-1-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-carbamicacid methyl ester;[2-(1,1-dioxo-1λ⁶-thiomorpholin-4-yl)ethyl]-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-carbamicacid methyl ester;(1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}propyl)-carbamicacid methyl ester;((S)-1,1-dioxo-tetrahydro-1λ⁶-thiophen-3-yl)-(2-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}ethyl)-carbamicacid methyl ester; 1-isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylicacid {(1S,3R,5R)-8-[3-(methyl-{2-[4-(tetrahydrofuran-2-carbonyl)piperazin-1-yl]ethyl}sulfamoyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid((1S,3R,5R)-8-{3-[4-(tetrahydrofuran-2-carbonyl)piperazine-1-sulfonyl]propyl}-8-azabicyclo-[3.2.1]oct-3-yl)amide;1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid{(1S,3R,5R)-8-[3-(4-acetylpiperazine-1-sulfonyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide;4-methanesulfonyl-piperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}propylester; 4-(tetrahydrofuran-2-carbonyl)piperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}propylester; 4-acetyl-piperazine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}propylester; and 4-hydroxypiperidine-1-carboxylic acid3-{(1S,3R,5R)-3-[(1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carbonyl)amino]-8-azabicyclo[3.2.1]-8-yl}-propylester; and pharmaceutically-acceptable salts thereof.
 6. The method ofclaim 1, wherein the disorder of reduced motility is chronicconstipation, irritable bowel syndrome, gastroparesis, post-operativeileus, intestinal pseudo-obstruction, drug-induced delayed transit, orfunctional dyspepsia.
 7. The method of claim 6, wherein the disorder ofreduced motility is chronic constipation, constipation-predominantirritable bowel syndrome, gastroparesis, or functional dyspepsia.
 8. Themethod of claim 5, wherein the disorder of reduced motility is chronicconstipation, irritable bowel syndrome, gastroparesis, post-operativeileus, intestinal pseudo-obstruction, drug-induced delayed transit, orfunctional dyspepsia.
 9. The method of claim 8, wherein the disorder ofreduced motility is chronic constipation, constipation-predominantirritable bowel syndrome, gastroparesis, or functional dyspepsia.
 10. Amethod of treating a mammal having a medical condition ameliorated bytreatment with a 5-hydroxytryptamine 4 (5-HT₄) receptor agonist, themethod comprising administering to the mammal, a therapeuticallyeffective amount of a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of formula (I) asdefined in claim 1 or a pharmaceutically-acceptable salt or stereoisomerthereof.
 11. The method of claim 10 wherein the medical condition isirritable bowel syndrome, chronic constipation, functional dyspepsia,delayed gastric emptying, gastroesophageal reflux disease,gastroparesis, post-operative ileus, intestinal pseudo-obstruction, ordrug-induced delayed transit.